U.S. patent application number 14/523481 was filed with the patent office on 2015-09-24 for 5h-cyclopenta[d]pyrimidines as akt protein kinase inhibitors.
The applicant listed for this patent is Array BioPharma Inc., Genentech, Inc.. Invention is credited to Josef R. Bencsik, James F. Blake, Martin F. Hentemann, Nicholas C. Kallan, Anna Leivers, Jun Liang, Ian S. Mitchell, Stephen T. Schlachter, Tony P. Tang, Eli M. Wallace, Rui Xu.
Application Number | 20150266852 14/523481 |
Document ID | / |
Family ID | 40578062 |
Filed Date | 2015-09-24 |
United States Patent
Application |
20150266852 |
Kind Code |
A1 |
Leivers; Anna ; et
al. |
September 24, 2015 |
5H-CYCLOPENTA[d]PYRIMIDINES AS AKT PROTEIN KINASE INHIBITORS
Abstract
Compounds of Formula I are useful for inhibiting AKT protein
kinases. Methods of using compounds of Formula I and stereoisomers
and pharmaceutically acceptable salts thereof, for in vitro, in
situ, and in vivo diagnosis, prevention or treatment of such
disorders in mammalian cells, or associated pathological conditions
are disclosed. ##STR00001##
Inventors: |
Leivers; Anna; (Boulder,
CO) ; Bencsik; Josef R.; (Boulder, CO) ;
Blake; James F.; (Boulder, CO) ; Hentemann; Martin
F.; (Boulder, CO) ; Kallan; Nicholas C.;
(Boulder, CO) ; Liang; Jun; (South San Francisco,
CA) ; Mitchell; Ian S.; (South San Francisco, CA)
; Schlachter; Stephen T.; (Boulder, CO) ; Wallace;
Eli M.; (Boulder, CO) ; Xu; Rui; (Boulder,
CO) ; Tang; Tony P.; (Boulder, CO) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Array BioPharma Inc.
Genentech, Inc. |
Boulder
South San Francisco |
CO
CA |
US
US |
|
|
Family ID: |
40578062 |
Appl. No.: |
14/523481 |
Filed: |
October 24, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
13678395 |
Nov 15, 2012 |
8895566 |
|
|
14523481 |
|
|
|
|
12812377 |
Jun 16, 2011 |
8329709 |
|
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PCT/US2009/030617 |
Jan 9, 2009 |
|
|
|
13678395 |
|
|
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|
61020129 |
Jan 9, 2008 |
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Current U.S.
Class: |
544/253 |
Current CPC
Class: |
A61P 25/00 20180101;
C07D 417/04 20130101; A61P 25/06 20180101; C07D 417/10 20130101;
C07D 487/04 20130101; C07D 239/70 20130101; A61P 9/00 20180101;
C07D 211/08 20130101; C07D 401/12 20130101; C07D 401/10 20130101;
C07D 409/04 20130101; C07D 403/04 20130101; C07D 211/46 20130101;
A61P 35/02 20180101; A61P 35/00 20180101; C07D 403/10 20130101;
A61P 43/00 20180101; C07D 401/04 20130101 |
International
Class: |
C07D 401/10 20060101
C07D401/10; C07D 211/08 20060101 C07D211/08; C07D 211/46 20060101
C07D211/46 |
Claims
1-42. (canceled)
43. A process for preparing a compound of formula 40: ##STR00084##
or a salt thereof, comprising converting a corresponding compound
of formula 37: ##STR00085## or a salt thereof, to provide the
compound of formula 40 or a salt thereof, wherein: PG is an amine
protecting group; Hal is Br, Cl or I; R.sup.1 and R.sup.1a are
independently selected from hydrogen, methyl, ethyl,
--CH.dbd.CH.sub.2, --CH.sub.2OOH, CF.sub.3, CHF.sub.2 and
CH.sub.2F; R.sup.2 is selected from hydrogen, OH, OCH.sub.3 and F;
R.sup.2a is selected from hydrogen, methyl and F, or R.sup.2 and
R.sup.2a are oxo; G is phenyl optionally substituted with one to
four R.sup.a groups or a 5-6 membered heteroaryl optionally
substituted by a halogen; R.sup.5 is selected from hydrogen and
C.sub.1-C.sub.4 alkyl; and each R.sup.a is independently halogen,
C.sub.1-C.sub.6-alkyl, C.sub.3-C.sub.6-cycloalkyl,
--O--(C.sub.1-C.sub.6-alkyl), CF.sub.3, --OCF.sub.3,
S(C.sub.1-C.sub.6-alkyl), CN, --OCH.sub.2-phenyl, NH.sub.2,
--NO.sub.2, --NH--(C.sub.1-C.sub.6-alkyl),
--N--(C.sub.1-C.sub.6-alkyl).sub.2, piperidine, pyrrolidine,
CH.sub.2F, CHF.sub.2, --OCH.sub.2F, --OCHF.sub.2, --OH,
--SO.sub.2(C.sub.1-C.sub.6-alkyl), C(O)NH.sub.2,
C(O)NH(C.sub.1-C.sub.6-alkyl) or
C(O)N(C.sub.1-C.sub.6-alkyl).sub.2.
44. The process of claim 43, wherein the compound of formula 40 or
a salt thereof is a compound of formula 39: ##STR00086## or a salt
thereof.
45. The process of claim 43, wherein the compound of formula 37 or
a salt thereof is first converted to an organometallic before being
converted to the compound of formula 40 or a salt thereof.
46. The process of claim 43, further comprising deprotection and
optional functionalization.
Description
PRIORITY OF INVENTION
[0001] This application is a continuation of U.S. patent
application Ser. No. 13/678,395, filed on Nov. 15, 2012, which is a
continuation of U.S. patent application Ser. No. 12/812,377, now
U.S. Pat. No. 8,329,709, which is a 35 U.S.C. 371 national stage
application of International Patent Application No.
PCT/US2009/030617, filed Jan. 9, 2009, and claims priority to U.S.
Provisional Application No. 61/020,129 that was filed on Jan. 9,
2008, which are incorporated herein by reference in their
entirety.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] The present invention relates to novel inhibitors of
serine/threonine protein kinases (e.g., AKT and related kinases),
to pharmaceutical compositions comprising the compounds, to a
process for making the compounds and to the use of the compounds in
therapy. More particularly it relates to certain 4-substituted
5H-cyclopenta[d]pyrimidines useful in the treatment and prevention
of hyperproliferative diseases, such as cancer and inflammation, in
mammals.
[0004] 2. Description of the State of the Art
[0005] Protein kinases (PK) are enzymes that catalyze the
phosphorylation of hydroxy groups on tyrosine, serine and threonine
residues of proteins by transfer of the terminal (gamma) phosphate
from ATP. Through signal transduction pathways, these enzymes
modulate cell growth, differentiation and proliferation, i.e.,
virtually all aspects of cell life in one way or another depend on
PK activity (Hardie, G. and Hanks, S. (1995) The Protein Kinase
Facts Book I and II, Academic Press, San Diego, Calif.).
Furthermore, abnormal PK activity has been related to a host of
disorders, ranging from relatively non-life threatening diseases
such as psoriasis to extremely virulent diseases such as
glioblastoma (brain cancer). Protein kinases are an important
target class for therapeutic modulation (Cohen, P. (2002) Nature
Rev. Drug Discovery 1:309).
[0006] Significantly, atypical protein phosphorylation and/or
expression is often reported to be one of the causative effects of
abnormal cellular proliferation, metastasis and cell survival in
cancer. The abnormal regulation and/or expression of various
kinases, including Akt, VEGF, ILK, ROCK, p70S6K, Bcl, PKA, PKC,
Raf, Src, PDK1, ErbB2, MEK, IKK, Cdk, EGFR, BAD, CHK1, CHK2 and
GSK3 amongst numerous others, has been specifically implicated in
cancer.
[0007] Protein kinases include two classes; protein tyrosine
kinases (PTK) and serine-threonine kinases (STK). The Protein
Kinase B/Akt enzymes are a group of serine/threonine kinases that
are overexpressed in a variety of human tumors. One of the
best-characterized targets of the PI3K lipid products is the 57 KD
serine/threonine protein kinase Akt, downstream of PI3K in the
signal transduction pathway (Hemmings, B. A. (1997) Science
275:628; Hay N. (2005) Cancer Cell 8:179-183). Akt is the human
homologue of the protooncogene v-akt of the acutely transforming
retrovirus AKT8. Due to its high sequence homology to protein
kinases A and C, Akt is also called Protein Kinase B (PKB) and
Related to A and C (RAC). Three isoforms of Akt are known to exist,
namely Akt1, Akt2 and Akt3, which exhibit an overall homology of
80% (Staal, S. P. (1987) Proc. Natl. Acad. Sci. 84:5034; Nakatani,
K. (1999) Biochem. Biophys. Res. Commun. 257:906; Li et al (2002)
Current Topics in Med. Chem. 2:939-971; WO 2005/113762). The Akt
isoforms share a common domain organization that consists of a
pleckstrin homology domain at the N-terminus, a kinase catalytic
domain, and a short regulatory region at the C-terminus. In
addition, both Akt2 and Akt3 exhibit splice variants. Upon
recruitment to the cell membrane by PtdInd(3,4,5)P.sub.3, Akt is
phosphorylated (activated) by PDK1 at T308, T309 and T305 for
isoforms Akt1 (PKB.alpha.), Akt2 (PKB.beta.) and Akt3 (PKB.gamma.),
respectively, and at S473, S474 and S472 for isoforms Akt1, Akt2
and Akt3, respectively. Such phosphorylation is believed to occur
by the mTOR-Rictor complex, although PDK1 (Balendran, A., (1999)
Curr. Biol. 9:393), autophosphorylation (Toker, A. (2000) J. Biol.
Chem. 275:8271) and integrin-linked kinase (ILK) (Delcommenne, M.
(1998) Proc. Natl. Acad. Sci. USA, 95:11211) have been implicated
in this process. Akt activation requires its phosphorylation on
residue Ser 473 in the C-terminal hydrophobic motif (Brodbeck et al
(1999) J. Biol. Chem. 274:9133-9136; Coffer et al (1991) Eur. J.
Biochem. 201:475-481; Alessi et al (1997) Curr. Biol. 7:261-269).
Although monophosphorylation of Akt activates the kinase,
bis(phosphorylation) is required for maximal kinase activity.
[0008] Akt is believed to assert its effect on cancer by
suppressing apoptosis and enhancing both angiogenesis and
proliferation (Toker et al (2006) Cancer Res. 66(8):3963-3966). Akt
is overexpressed in many forms of human cancer including, but not
limited to, colon (Zinda et al (2001) Clin. Cancer Res. 7:2475),
ovarian (Cheng et al (1992) Proc. Natl. Acad. Sci. USA 89:9267),
brain (Haas Kogan et al (1998) Curr. Biol. 8:1195), lung (Brognard
et al (2001) Cancer Res. 61:3986), pancreatic (Bellacosa et al
(1995) Int. J. Cancer 64:280-285; Cheng et al (1996) Proc. Natl.
Acad. Sci. 93:3636-3641), prostate (Graff et al (2000) J. Biol.
Chem. 275:24500) and gastric carcinomas (Staal et al (1987) Proc.
Natl. Acad. Sci. USA 84:5034-5037).
[0009] The PI3K/Akt/mammalian target of rapamycin (mTOR) pathway
has been explored for targeted small molecule inhibitor therapy
(Georgakis, G. and Younes, A. (2006) Expert Rev. Anticancer Ther.
6(1):131-140; Granville et al (2006) Clin. Cancer Res.
12(3):679-689) Inhibition of PI3K/Akt signaling induces apoptosis
and inhibits the growth of tumor cells that have elevated Akt
levels (Kim et al (2005) Current Opinion in Investig. Drugs
6(12):1250-1258; Luo et al (2005) Molecular Cancer Ther.
4(6):977-986).
[0010] The development of kinase inhibitors that target abnormally
regulated pathways and ultimately result in disease is of enormous
ethical and commercial interest to the medical and pharmaceutical
community. A compound that inhibits (1) recruitment of Akt to the
cell membrane, (2) activation by PDK1 or PDK2, (3) substrate
phosphorylation, or (4) one of the downstream targets of Akt could
be a valuable anticancer agent, either as a stand-alone therapy or
in conjunction with other accepted procedures.
[0011] United States Patent Application Publication 2005/0130954
discloses inter alia, a variety of compounds that act as AKT
inhibitors. The compounds are said to be useful in the treatment of
hyperproliferative diseases such as cancer.
SUMMARY OF THE INVENTION
[0012] In one aspect, the invention relates to compounds that are
inihibitors of AKT protein kinases. Accordingly, the compounds of
the invention are useful in the treatment of hyperproliferative
diseases, such as cancer and inflammation, in mammals.
[0013] More specifically, one aspect of the present invention
provides compounds of Formula I:
##STR00002##
and stereoisomers and pharmaceutically acceptable salts thereof,
wherein R.sup.1, R.sup.1a, R.sup.2, R.sup.2a, L and A are as
defined herein.
[0014] Another aspect of the present invention provides methods of
preventing or treating a disease or disorder modulated by AKT,
comprising administering to a mammal in need of such treatment an
effective amount of a compound of this invention or a stereoisomer
or pharmaceutically acceptable salt thereof. Examples of such
diseases and disorders include, but are not limited to,
hyperproliferative disorders (such as cancer), neurodegeneration,
cardiac hypertrophy, pain, migraine and neurotraumatic disease.
[0015] Another aspect of the present invention provides methods of
preventing or treating cancer, comprising administering to a mammal
in need of such treatment an effective amount of a compound of this
invention, or a stereoisomer or pharmaceutically acceptable salt
thereof, alone or in combination with one or more additional
compounds having anticancer properties.
[0016] In another aspect, the present invention provides a method
of inhibiting the production of AKT protein kinases in a mammal,
which comprises administering to said mammal a compound of Formula
I, or a stereoisomer or pharmaceutically acceptable salt thereof in
an amount effective to inhibit production of an AKT protein
kinase.
[0017] In still yet another aspect, the present invention provides
methods of inhibiting the activity of AKT protein kinases,
comprising contacting said kinase with a compound of Formula I.
[0018] The inventive compounds may be used advantageously in
combination with other known therapeutic agents. Accordingly, this
invention also provides pharmaceutical compositions comprising a
compound of Formula I or a stereoisomer or pharmaceutically
acceptable salt thereof, in combination with a second therapeutic
agent.
[0019] Another aspect of the present invention provides a method of
treating a hyperproliferative disease in a mammal comprising
administering a therapeutically effective amount of a compound of
this invention to the mammal.
[0020] Another aspect of the present invention provides the use of
a compound of this invention in the manufacture of a medicament for
the treatment of a hyperproliferative disease.
[0021] Another aspect of the present invention provides compounds
of the present invention for use in the treatment of
hyperproliferative diseases.
[0022] An additional aspect of the invention is the use of a
compound of Formula I, or a stereoisomer or pharmaceutically
acceptable salt thereof, for therapy. In one embodiment, the
therapy comprises the treatment of an AKT protein kinase-mediated
condition.
[0023] Another aspect of the present invention provides a
pharmaceutical composition comprising a compound of the present
invention in the treatment of a hyperproliferative disease.
[0024] Another aspect of the present invention provides a
pharmaceutical composition comprising a compound of the present
invention in the treatment of cancer.
[0025] Another aspect of the present invention provides a
pharmaceutical composition comprising a compound of this invention
or a pharmaceutically acceptable salt thereof, and a
pharmaceutically acceptable carrier or excipient.
[0026] Another aspect of the present invention includes methods of
preparing, methods of separation, and methods of purification of
the compounds of this invention.
DETAILED DESCRIPTION OF THE INVENTION
[0027] Reference will now be made in detail to certain embodiments
of the invention, examples of which are illustrated in the
accompanying structures and formulas. While the invention will be
described in conjunction with the enumerated embodiments, it will
be understood that they are not intended to limit the invention to
those embodiments. On the contrary, the invention is intended to
cover all alternatives, modifications, and equivalents, which may
be included within the scope of the present invention as defined by
the claims. One skilled in the art will recognize many methods and
materials similar or equivalent to those described herein, which
could be used in the practice of the present invention. The present
invention is in no way limited to the methods and materials
described. In the event that one or more of the incorporated
literature and similar materials differs from or contradicts this
application, including but not limited to defined terms, term
usage, described techniques, or the like, this application
controls.
DEFINITIONS
[0028] The term "alkyl" includes linear or branched-chain radicals
of carbon atoms. Some alkyl moieties have been abbreviated, for
example, methyl ("Me"), ethyl ("Et"), propyl ("Pr") and butyl
("Bu"), and further abbreviations are used to designate specific
isomers of compounds, for example, 1-propyl or n-propyl ("n-Pr"),
2-propyl or isopropyl ("i-Pr"), 1-butyl or n-butyl ("n-Bu"),
2-methyl-1-propyl or isobutyl ("i-Bu"), 1-methylpropyl or s-butyl
("s-Bu"), 1,1-dimethylethyl or t-butyl ("t-Bu") and the like. The
abbreviations are sometimes used in conjunction with elemental
abbreviations and chemical structures, for example, methanol
("MeOH") or ethanol ("EtOH").
[0029] Additional abbreviations used throughout the application
include, for example, benzyl ("Bz") and phenyl ("Ph").
[0030] The terms "treat" or "treatment" refer to therapeutic,
prophylactic, palliative or preventative measures. For purposes of
this invention, beneficial or desired clinical results include, but
are not limited to, alleviation of symptoms, diminishment of extent
of disease, stabilized (i.e., not worsening) state of disease,
delay or slowing of disease progression, amelioration or palliation
of the disease state, and remission (whether partial or total),
whether detectable or undetectable. "Treatment" can also mean
prolonging survival as compared to expected survival if not
receiving treatment. Those in need of treatment include those
already with the condition or disorder as well as those prone to
have the condition or disorder or those in which the condition or
disorder is to be prevented.
[0031] The phrases "therapeutically effective amount" or "effective
amount" mean an amount of a compound of the present invention that,
when administered to a mammal in need of such treatment, sufficient
to (i) treat or prevent the particular disease, condition, or
disorder, (ii) attenuate, ameliorate, or eliminate one or more
symptoms of the particular disease, condition, or disorder, or
(iii) prevent or delay the onset of one or more symptoms of the
particular disease, condition, or disorder described herein. The
amount of a compound that will correspond to such an amount will
vary depending upon factors such as the particular compound,
disease condition and its severity, the identity (e.g., weight) of
the mammal in need of treatment, but can nevertheless be routinely
determined by one skilled in the art.
[0032] The terms "cancer" and "cancerous" refer to or describe the
physiological condition in mammals that is typically characterized
by unregulated cell growth. A "tumor" comprises one or more
cancerous cells. Examples of cancer include, but are not limited
to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or
lymphoid malignancies. More particular examples of such cancers
include squamous cell cancer (e.g., epithelial squamous cell
cancer), lung cancer including small-cell lung cancer, non-small
cell lung cancer ("NSCLC"), adenocarcinoma of the lung and squamous
carcinoma of the lung, cancer of the peritoneum, hepatocellular
cancer, gastric or stomach cancer including gastrointestinal
cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian
cancer, liver cancer, bladder cancer, hepatoma, breast cancer,
colon cancer, rectal cancer, colorectal cancer, endometrial or
uterine carcinoma, salivary gland carcinoma, kidney or renal
cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic
carcinoma, anal carcinoma, penile carcinoma, skin cancer, including
melanoma, as well as head and neck cancer.
[0033] The phrase "pharmaceutically acceptable" indicates that the
substance or composition must be compatible chemically and/or
toxicologically, with the other ingredients comprising a
formulation, and/or the mammal being treated therewith.
[0034] The phrase "pharmaceutically acceptable salt," as used
herein, refers to pharmaceutically acceptable organic or inorganic
salts of a compound of the invention.
[0035] The compounds of this invention also include other salts of
such compounds which are not necessarily pharmaceutically
acceptable salts, and which may be useful as intermediates for
preparing and/or purifying compounds of this invention and/or for
separating enantiomers of compounds of this invention.
[0036] The term "mammal" means a warm-blooded animal that has or is
at risk of developing a disease described herein and includes, but
is not limited to, guinea pigs, dogs, cats, rats, mice, hamsters,
and primates, including humans.
AKT Inhibitors
[0037] The present invention provides compounds, and pharmaceutical
formulations thereof, that are potentially useful in the treatment
of diseases, conditions and/or disorders modulated by AKT.
[0038] One embodiment of this invention provides compounds of
Formula I:
##STR00003##
[0039] and stereoisomers and pharmaceutically acceptable salts
thereof, wherein:
[0040] R.sup.1 and R.sup.1a are independently selected from
hydrogen, methyl, ethyl, --CH.dbd.CH.sub.2, --CH.sub.2OH, CF.sub.3,
CHF.sub.2 or CH.sub.2F;
[0041] R.sup.2 is selected from hydrogen, OH, OCH.sub.3 or F;
[0042] R.sup.2a is selected from hydrogen, methyl or F, or
[0043] R.sup.2 and R.sup.2a are oxo;
[0044] L is selected from:
##STR00004##
wherein the single wavy line is where L attaches to A and the
double wavy line is where L attaches to the pyrimidine;
[0045] A is:
##STR00005##
[0046] X is a direct bond from L to Y, CH.sub.2, O, C.dbd.O, NH or
C(.dbd.O)NH;
[0047] Y is CH or N;
[0048] Z is absent, CH.sub.2 or 0, wherein L, X, Y, Z and b are
selected so that any nitrogen is not bonded directly to another
nitrogen;
[0049] G is phenyl optionally substituted with one to four R.sup.a
groups or a 5-6 membered heteroaryl optionally substituted by a
halogen;
[0050] R.sup.3 and R.sup.4 are independently selected from hydrogen
or methyl;
[0051] R.sup.5 and R.sup.6 are independently selected from hydrogen
or C.sub.1-C.sub.4 alkyl;
[0052] a is 0 or 1;
[0053] b is 0, 1 or 2; and
[0054] each R.sup.a is independently halogen,
C.sub.1-C.sub.6-alkyl, C.sub.3-C.sub.6-cycloalkyl,
--O--(C.sub.1-C.sub.6-alkyl), CF.sub.3, --OCF.sub.3,
S(C.sub.1-C.sub.6-alkyl), CN, --OCH.sub.2-phenyl, NH.sub.2,
--NO.sub.2, --NH--(C.sub.1-C.sub.6-alkyl),
--N--(C.sub.1-C.sub.6-alkyl).sub.2, piperidine, pyrrolidine,
CH.sub.2F, CHF.sub.2, --OCH.sub.2F, --OCHF.sub.2, --OH,
--SO.sub.2(C.sub.1-C.sub.6-alkyl), C(O)NH.sub.2,
C(O)NH(C.sub.1-C.sub.6-alkyl), and
C(O)N(C.sub.1-C.sub.6-alkyl).sub.2; or
[0055] b is 1, R.sup.3 is hydrogen and R.sup.4 and R.sup.5 together
with the atoms to which they are attached form an optionally
substituted 5-6 membered heterocyclic ring having one ring nitrogen
atom, and R.sup.6 is selected from the group consisting of hydrogen
or C.sub.1-C.sub.4 alkyl optionally substituted with OH or
O(C.sub.1-C.sub.3 alkyl), such that A has the structure:
##STR00006##
[0056] R.sup.c and R.sup.d are independently selected from hydrogen
and methyl; and
[0057] c is 1 or 2; or
[0058] b is 1, Z is CH.sub.2 and R.sup.5 and Y together with the
atoms to which they are attached form an optionally substituted 6
membered heterocyclic ring having one ring nitrogen atom, and
R.sup.6 is selected from the group consisting of hydrogen or
C.sub.1-C.sub.4 alkyl optionally substituted with OH or
O(C.sub.1-C.sub.3 alkyl), such that A has the structure:
##STR00007##
[0059] In certain embodiments, R.sup.1 is methyl.
[0060] In certain embodiments, R.sup.1a is hydrogen.
[0061] In certain embodiments, R.sup.2 is hydrogen.
[0062] In certain embodiments, R.sup.2 is OH.
[0063] In certain embodiments, R.sup.2a is hydrogen.
[0064] In certain embodiments, Formula I is selected from:
##STR00008##
[0065] In certain embodiments, Formula I is selected from:
##STR00009##
[0066] In certain embodiments, L is selected from:
##STR00010##
[0067] In certain embodiments, L is:
##STR00011##
wherein the single wavy line is where L attaches A and the double
wavy line is where L attaches to the pyrimidine, providing
compounds of Formula Ia having the structure:
##STR00012##
[0068] In certain embodiments, L is:
##STR00013##
wherein the single wavy line is where L attaches A and the double
wavy line is where L attaches to the pyrimidine, providing
compounds of Formula Ib having the structure:
##STR00014##
[0069] In certain embodiments, L is:
##STR00015##
wherein the single wavy line is where L attaches A and the double
wavy line is where L attaches to the pyrimidine, providing
compounds of Formula Ic having the structure:
##STR00016##
[0070] In certain embodiments, L is:
##STR00017##
wherein the single wavy line is where L attaches A and the double
wavy line is where L attaches to the pyrimidine, providing
compounds of Formula Id having the structure:
##STR00018##
[0071] In certain embodiments, L is:
##STR00019##
wherein the single wavy line is where L attaches A and the double
wavy line is where L attaches to the pyrimidine, providing
compounds of Formula Ie having the structure:
##STR00020##
[0072] In certain embodiments, L is:
##STR00021##
wherein the single wavy line is where L attaches A and the double
wavy line is where L attaches to the pyrimidine, providing
compounds of Formula If having the structure:
##STR00022##
[0073] In certain embodiments, L is:
##STR00023##
wherein the single wavy line is where L attaches A and the double
wavy line is where L attaches to the pyrimidine, providing
compounds of Formula Ig having the structure:
##STR00024##
[0074] In certain embodiments, L is:
##STR00025##
wherein the single wavy line is where L attaches A and the double
wavy line is where L attaches to the pyrimidine, providing
compounds of Formula Ih having the structure:
##STR00026##
[0075] In certain embodiments, L is:
##STR00027##
wherein the single wavy line is where L attaches A and the double
wavy line is where L attaches to the pyrimidine, providing
compounds of Formula Ii having the structure:
##STR00028##
[0076] In certain embodiments, A is:
##STR00029##
[0077] In certain embodiments, X is a direct bond from L to Y,
CH.sub.2, O, C.dbd.O, NH or C(.dbd.O)NH.
[0078] In certain embodiments, X is a direct bond from L to Y.
[0079] In certain embodiments, X is CH.sub.2.
[0080] In certain embodiments, X is O.
[0081] In certain embodiments, X is C.dbd.O.
[0082] In certain embodiments, X is NH.
[0083] In certain embodiments, X is C(.dbd.O)NH.
[0084] In certain embodiments, Y is CH or N.
[0085] In certain embodiments, Y is N.
[0086] In certain embodiments, Y is CH.
[0087] In certain embodiments, Z is absent, CH.sub.2 or O.
[0088] In certain embodiments, Z is absent or O.
[0089] In certain embodiments, Z is absent.
[0090] In certain embodiments, Z is CH.sub.2.
[0091] In certain embodiments, Z is O.
[0092] The compounds of Formula I are such that L, X, Y, Z and b
are selected so that any nitrogen is not bonded directly to another
nitrogen.
[0093] In certain embodiments, X is a direct bond from L to Y, Y is
CH.sub.2 and Z is O, such that A has the structure A1:
##STR00030##
[0094] In certain embodiments of A1, a is 0.
[0095] In certain embodiments of A1, b is 2.
[0096] In certain embodiments of A1, R.sup.3 is hydrogen.
[0097] In certain embodiments of A1, R.sup.4 is hydrogen.
[0098] In certain embodiments of A1, R.sup.5 is C.sub.1-C.sub.4
alkyl. In certain embodiments of A1, R.sup.5 is methyl.
[0099] In certain embodiments of A1, R.sup.6 is C.sub.1-C.sub.4
alkyl. In certain embodiments of A1, R.sup.6 is methyl.
[0100] In certain embodiments, X is C(.dbd.O)NH, Y is CH and Z is
absent, such that A has the structure A2:
##STR00031##
[0101] In certain embodiments of A2, a is 1.
[0102] In certain embodiments of A2, b is 1.
[0103] In certain embodiments of A2, R.sup.3 is hydrogen.
[0104] In certain embodiments of A2, R.sup.4 is hydrogen.
[0105] In certain embodiments of A2, R.sup.5 is hydrogen.
[0106] In certain embodiments of A2, R.sup.6 is hydrogen.
[0107] In certain embodiments, X is a direct bond from L to Y, Y is
CH and Z is absent, such that A has the structure A3:
##STR00032##
[0108] In certain embodiments of A3, a is 0.
[0109] In certain embodiments of A3, b is 1.
[0110] In certain embodiments of A3, R.sup.3 is hydrogen.
[0111] In certain embodiments of A3, R.sup.4 is hydrogen.
[0112] In certain embodiments of A3, R.sup.5 is C.sub.1-C.sub.4
alkyl. In certain embodiments of A3, R.sup.5 is isopropyl.
[0113] In certain embodiments of A3, R.sup.6 is hydrogen.
[0114] In certain embodiments, X is a direct bond from L to Y, Y is
CH and Z is absent, such that A has the structure A4:
##STR00033##
[0115] In certain embodiments of A4, a is 0.
[0116] In certain embodiments of A4, a is 1.
[0117] In certain embodiments of A4, b is 0.
[0118] In certain embodiments of A4, b is 1.
[0119] In certain embodiments of A4, R.sup.3 is hydrogen.
[0120] In certain embodiments of A4, R.sup.4 is hydrogen.
[0121] In certain embodiments of A4, R.sup.5 is hydrogen.
[0122] In certain embodiments of A4, R.sup.5 is C.sub.1-C.sub.4
alkyl. In certain embodiments of A4, R.sup.5 is isopropyl.
[0123] In certain embodiments of A4, R.sup.6 is hydrogen.
[0124] In certain embodiments, X is NH, Y is CH and Z is absent,
such that A has the structure A5:
##STR00034##
[0125] In certain embodiments of A5, a is 0.
[0126] In certain embodiments of A5, b is 1.
[0127] In certain embodiments of A5, R.sup.3 is hydrogen.
[0128] In certain embodiments of A5, R.sup.4 is hydrogen.
[0129] In certain embodiments of A5, R.sup.5 is C.sub.1-C.sub.4
alkyl. In certain embodiments of A5, R.sup.5 is isopropyl.
[0130] In certain embodiments of A5, R.sup.6 is hydrogen.
[0131] In certain embodiments, X is C.dbd.O, Y is N and Z is
absent, such that A has the structure A6:
##STR00035##
[0132] In the embodiments of A6, b must be 1 or 2.
[0133] In certain embodiments of A6, a is 1.
[0134] In certain embodiments of A6, b is 2.
[0135] In certain embodiments of A6, R.sup.3 is hydrogen.
[0136] In certain embodiments of A6, R.sup.4 is hydrogen.
[0137] In certain embodiments of A6, R.sup.5 is hydrogen.
[0138] In certain embodiments of A6, R.sup.6 is hydrogen.
[0139] In certain embodiments, X is C.dbd.O, Y is CH and Z is
absent, such that A has the structure A7:
##STR00036##
[0140] In certain embodiments of A7, a is 0.
[0141] In certain embodiments of A7, b is 1.
[0142] In certain embodiments of A7, R.sup.3 is hydrogen.
[0143] In certain embodiments of A7, R.sup.4 is hydrogen.
[0144] In certain embodiments of A7, R.sup.5 is C.sub.1-C.sub.4
alkyl. In certain embodiments of A7, R.sup.5 is tert-butyl. In
certain embodiments of A7, R.sup.5 is isopropyl.
[0145] In certain embodiments of A7, R.sup.6 is hydrogen.
[0146] In certain embodiments, R.sup.3 is hydrogen.
[0147] In certain embodiments, R.sup.3 is methyl.
[0148] In certain embodiments, R.sup.4 is hydrogen.
[0149] In certain embodiments, R.sup.4 is methy.
[0150] In certain embodiments, R.sup.5 is hydrogen.
[0151] In certain embodiments, R.sup.5 is C.sub.1-C.sub.4
alkyl.
[0152] In certain embodiments, R.sup.5 is methyl.
[0153] In certain embodiments, R.sup.5 is isopropyl.
[0154] In certain embodiments, R.sup.5 is tert-butyl.
[0155] In certain embodiments, R.sup.6 is hydrogen.
[0156] In certain embodiments, R.sup.6 is methyl.
[0157] In certain embodiments, a is 1.
[0158] In certain embodiments, a is 0.
[0159] In certain embodiments, b is 0.
[0160] In certain embodiments, b is 1.
[0161] In certain embodiments, b is 2.
[0162] In certain embodiments, Z is O and b is 2.
[0163] In certain embodiments, b is 1, R.sup.3 is hydrogen and
R.sup.4 and R.sup.5 together with the atoms to which they are
attached form an optionally substituted 5-6 membered heterocyclic
ring having one ring nitrogen atom, such that A has the structure
A8:
##STR00037##
wherein c is 1 or 2, R.sup.c and R.sup.d are independently selected
from hydrogen and methyl, and R.sup.6 is selected from the group
consisting of H or C.sub.1-C.sub.4 alkyl optionally substituted
with OH or O(C.sub.1-C.sub.3 alkyl).
[0164] In certain embodiments, c is 1, such that A has the
structure A8a:
##STR00038##
[0165] In certain embodiments of A8a, X is C.dbd.O.
[0166] In certain embodiments of A8a, Y is CH.
[0167] In certain embodiments of A8a, Z is absent.
[0168] In certain embodiments of A8a, a is 0.
[0169] In certain embodiments of A8a, R.sup.c is methyl.
[0170] In certain embodiments of A8a, R.sup.d is methyl.
[0171] In certain embodiments of A8a, R.sup.6 is hydrogen.
[0172] In certain embodiments, c is 2, such that A has the
structure A8b:
##STR00039##
[0173] In certain embodiments, b is 1, Z is CH.sub.2 and R.sup.5
and Y together with the atoms to which they are attached form an
optionally substituted 6 membered heterocyclic ring having one ring
nitrogen atom, and R.sup.6 is selected from the group consisting of
hydrogen or C.sub.1-C.sub.4 alkyl optionally substituted with OH or
O(C.sub.1-C.sub.3 alkyl), such that A has the structure A9:
##STR00040##
[0174] In certain embodiments of Formula A9, X is a direct bond
from L to Y.
[0175] In certain embodiments of Formula A9, a is 0.
[0176] In certain embodiments of Formula A9, R.sup.6 is
hydrogen.
[0177] In certain embodiments, G is phenyl optionally substituted
with one to four R.sup.a groups or a 5-6 membered heteroaryl
optionally substituted by a halogen.
[0178] In certain embodiments, each R.sup.a is independently
halogen, C.sub.1-C.sub.6-alkyl, C.sub.3-C.sub.6-cycloalkyl,
--O--(C.sub.1-C.sub.6-alkyl), CF.sub.3, --OCF.sub.3,
S(C.sub.1-C.sub.6-alkyl), CN, --OCH.sub.2-phenyl, NH.sub.2,
--NO.sub.2, --NH--(C.sub.1-C.sub.6-alkyl),
--N--(C.sub.1-C.sub.6-alkyl).sub.2, piperidine, pyrrolidine,
CH.sub.2F, CHF.sub.2, --OCH.sub.2F, --OCHF.sub.2, --OH,
--SO.sub.2(C.sub.1-C.sub.6-alkyl), C(O)NH.sub.2,
C(O)NH(C.sub.1-C.sub.6-alkyl), and
C(O)N(C.sub.1-C.sub.6-alkyl).sub.2.
[0179] Referring to the G group of Formula I, examples include
phenyl optionally substituted with one or more R.sup.a groups
independently selected from F, Cl, Br, I, methyl, ethyl, isopropyl,
tert-butyl, cyclopropyl, CN, CF.sub.3, -OMe, -OEt, --OCF.sub.3,
--NO.sub.2, -SMe and --OCH.sub.2Ph. Exemplary embodiments of G
include phenyl, 2-chlorophenyl, 3-chlorophenyl, 4-chlorophenyl,
4-fluorophenyl, 4-bromophenyl, 4-methylphenyl, 4-ethylphenyl,
4-isopropylphenyl, 4-trifluoromethylphenyl, 4-cyanophenyl,
4-methoxyphenyl, 4-ethoxyphenyl, 4-thiomethylphenyl,
4-trifluoromethoxyphenyl, 4-cyclopropylphenyl,
4-chloro-3-fluorophenyl, 3,4-difluorophenyl,
4-bromo-3-fluorophenyl, 3-fluoro-4-methylphenyl,
3-fluoro-4-methoxyphenyl, 3-fluoro-4-trifluoromethylphenyl,
4-cyano-3-fluorophenyl, 3,4-dichlorophenyl, 2,4-dichlorophenyl,
2,4-difluorophenyl, 2-chloro-4-fluorophenyl,
2-fluoro-4-chlorophenyl, 3,5-dichlorophenyl. 3,5-difluorophenyl,
3-chloro-5-fluorophenyl, 3-chloro-4-fluorophenyl,
3-bromo-4-fluorophenyl, 3,5-difluoro-4-chlorophenyl,
2,3-difluoro-4-chlorophenyl, 2,5-difluoro-4-chlorophenyl,
3,5-difluoro-4-bromophenyl, 2,3-difluoro-4-bromophenyl,
2,5-difluoro-4-bromophenyl, 4-(OCH.sub.2Ph)-phenyl, 4-chlorophenyl,
2,4-dichlorophenyl, 3,4-dichlorophenyl, 4-chloro-3-fluorophenyl,
3-chloro-4-fluorophenyl, 3-fluoro-4-bromophenyl, 4-fluorophenyl,
3,4-difluorophenyl, 2,4-difluorophenyl 4-bromophenyl,
4-chloro-2-fluorophenyl, 4-methoxyphenyl, 4-methylphenyl,
4-cyanophenyl, 4-trifluoromethylphenyl, 4-iodophenyl,
4-nitrophenyl, 4-tert-butylphenyl, 2-fluorophenyl,
3-trifluoromethylphenyl, 2-fluoro-4-trifluoromethylphenyl,
3-fluoro-4-trifluoromethoxyphenyl, 3-fluoro-4-trifluoromethylphenyl
and 4-trifluoromethoxyphenyl.
[0180] In certain embodiments, G is 4-chlorophenyl, 4-bromophenyl,
4-trifluoromethylphenyl or 2,4-dichlorophenyl.
[0181] Referring to the G group of Formula I, the phrase "5-6
membered heteroaryl optionally substituted by a halogen" includes
thiophenes and pyridines, optionally substituted by halogens.
Particular examples include, but are not limited to, the
structures:
##STR00041##
[0182] It will be appreciated that certain compounds of the
invention may contain asymmetric or chiral centers, and therefore
exist in different stereoisomeric forms. It is intended that all
stereoisomeric forms of the compounds of the invention, including
but not limited to, diastereomers, enantiomers and atropisomers, as
well as mixtures thereof such as racemic mixtures, form part of the
present invention.
[0183] In the structures shown herein, where the stereochemistry of
any particular chiral atom is not specified, then all stereoisomers
are contemplated and included as the compounds of the invention.
Where stereochemistry is specified by a solid wedge or dashed line
representing a particular configuration, then that stereoisomer is
so specified and defined.
[0184] It will be further appreciated that the compounds of the
present invention may exist in unsolvated as well as solvated forms
with pharmaceutically acceptable solvents such as water, ethanol,
and the like, and it is intended that the invention embrace both
solvated and unsolvated forms.
Synthesis of Compounds
[0185] Compounds of the present invention may be synthesized by
synthetic routes that include processes analogous to those
well-known in the chemical arts, particularly in light of the
description contained herein. The starting materials are generally
available from commercial sources such as Sigma-Aldrich (St. Louis,
Mo.), Alfa Aesar (Ward Hill, Mass.), or TCI (Portland, Oreg.), or
are readily prepared using methods well known to those skilled in
the art (e.g., prepared by methods generally described in Louis F.
Fieser and Mary Fieser, Reagents for Organic Synthesis, v. 1-19,
Wiley, N.Y. (1967-1999 ed.), or Beilsteins Handbuch der organischen
Chemie, 4, Aufl. ed. Springer-Verlag, Berlin, including supplements
(also available via the Beilstein online database)).
[0186] For illustrative purposes, Schemes 1-20 shows a general
method for preparing the compounds of the present invention as well
as key intermediates. For a more detailed description of the
individual reaction steps, see the Examples section below. Those
skilled in the art will appreciate that other synthetic routes may
be used to synthesize the inventive compounds. Although specific
starting materials and reagents are depicted in the Schemes and
discussed below, other starting materials and reagents can be
easily substituted to provide a variety of derivatives and/or
reaction conditions. In addition, many of the compounds prepared by
the methods described below can be further modified in light of
this disclosure using conventional chemistry well known to those
skilled in the art.
##STR00042##
[0187] Scheme 1 shows a method of preparing compounds 7, wherein
Hal is Br, Cl or I. According to Scheme 1, intermediate 3 can be
prepared by brominating (+)-pulegone 1 to provide the di-bromide 2.
The di-bromide 2 is then treated with a base, such as sodium
ethoxide. Oxidative cleavage (e.g., ozonolysis at -80.degree. C. to
-50.degree. C.) of the pulegenate 3, wherein R.sup.f is
C.sub.1-C.sub.3 alkyl, gives the ketoester 4. The pyrimidine ring 5
is constructed by reaction of the ketoester 4 with thiourea in the
presence of a base, such as KOH. The mercapto group at the
2-position of compound 5 is eliminated by reduction (e.g., Raney Ni
in ammonia) to give compound 6. Alternatively, the ketoester 4 can
be converted to the same hydroxypyrimidine 6 by treatment with, for
example, an ammonia synthon, such as NH.sub.4OAc, followed by
treatment with NH.sub.4CO.sub.2H in the presence of formamide at
50.degree. C. to 250.degree. C. and/or at high pressure. Activation
of the hydroxypyrimidine 6 (e.g., POCl.sub.3) provides the
4-halopyrimidine 7.
##STR00043##
[0188] Scheme 2 shows a method of preparing compound 11, wherein
Hal is Br, Cl or I and R.sup.1 and R.sup.1a are defined herein.
According to Scheme 2, amination of compound 8, wherein R.sup.g is
C.sub.1-C.sub.3 alkyl, using an ammonia synthon (e.g., NH.sub.4OAc)
gives compound 9. Pyrimidine formation using, for example, ammonium
formate in the presence of formamide at 50.degree. C. to
250.degree. C. and/or at high pressure gives the bicyclic unit 10.
Activation of compound 10 using, for example, POCl.sub.3 or
SOCl.sub.2 gives the activated pyrimidine 11.
##STR00044##
[0189] Scheme 3 shows an alternative method of preparing compound
11, wherein Hal is Br, Cl or I and R.sup.1 and R.sup.1a groups are
defined herein. According to Scheme 3, the pyrimidine ring is
constructed by reacting the ketoester 8, wherein R.sup.g is
C.sub.1-C.sub.3 alkyl, with thiourea in the presence of a base,
such as KOH. The mercapto group at 2-position of compound 12 is
eliminated by reduction (e.g., Raney Ni in ammonia) to give 10.
Activation of compound 10 using, for example, POCl.sub.3 or
SOCl.sub.2 gives the activated pyrimidine 11.
##STR00045##
[0190] Scheme 4 shows a method of preparing compounds 16, 17, 18
and 19, wherein Hal is Br, Cl or I and R.sup.1 and R.sup.1a are
defined herein. According to Scheme 4, oxidation of the
4-halopyrimidine 11 with an oxidizing agent, such as
m-chloroperbenzoic acid ("m-CPBA"), Oxone or hydrogen peroxide
provides the N-oxide 13. Rearrangement of the N-oxide 13 with
acetic anhydride yields the intermediate 14, wherein R.sup.j is
methyl if acetic anhydride is used. Compound 14 is then hydrolyzed
under mild conditions (e.g., LiOH in an aqueous/organic solvent
mixture at 0.degree. C. to room temperature) to give the alcohol
15. Compound 15 is then either: 1) subjected to separation (e.g.,
chromatography with a chiral or achiral stationary phase); 2)
functionalized (e.g., 4-nitrobenzoyl chloride, NEt.sub.3) to
facilitate separation, separated (e.g., chromatography or
recrystallisation) and then hydrolyzed upon treatment with a base,
such as lithium hydroxide, in an aqueous/organic solvent mixture at
0.degree. C. to room temperature; or 3) oxidized (e.g., Swern
oxidation) followed by an asymmetric reduction (for example, a
catalytic chiral catalyst in the presence of hydrogen, the
Corey-Bakshi-Shibata catalyst ("CBS catalyst") or a borohydride
reducing agent in the presence of a chiral ligand). All
alternatives provide a route into the separate diastereomers 16 and
17.
[0191] Optionally, the 7-hydroxy group of compounds 16 and 17 may
be alkylated with an alkylating reagent, such as an alkyl halide
(e.g., MeI), in the presence of a base, such as NaH or KOH, to
provide compounds 18 and 19.
##STR00046##
[0192] Scheme 5 shows a method of preparing compounds 20 and 21,
wherein R.sup.k is halogen or NO.sub.2, Hal is Br, Cl or I and
R.sup.1 and R.sup.1a are defined herein. According to Scheme 5,
oxidation of the 4-haloopyrimidine 11 with an oxidizing agent, such
as m-CPBA, Oxone or hydrogen peroxide, provides the N-oxide 13.
Rearrangement of the N-oxide 13 with acetic anhydride yields the
intermediate 14, wherein R.sup.j is methyl if acetic anhydride is
used. Compound 14 is then hydrolyzed under mild conditions (e.g.,
LiOH in an aqueous/organic solvent mixture at 0.degree. C. to room
temperature) to give the alcohol 15. Compound 15 is then
functionalized (e.g., 4-nitrobenzoyl chloride or 4-bromobenzoyl
chloride in the presence of NEt.sub.3 at -20.degree. C. to
50.degree. C.) and separated (e.g., chromatography or
recrystallization) to provide a route into the separate, protected
diastereomers 20 and 21.
##STR00047##
[0193] Scheme 6 shows a method for the formation of compounds 26
and 28, wherein R.sup.1, R.sup.1a, R.sup.2 and R.sup.2a are as
defined herein. A Pd-mediated coupling between the halopyrimidine
22, wherein Hal is Br, Cl or I and R.sup.k is halogen or NO.sub.2,
and an appropriately substituted boronic acid or ester 23, wherein
PG is an amine protecting group and R.sup.m is hydrogen or alkyl
optionally substituted with OH or the two R.sup.m groups together
with the atoms to which they are attached form a 5 or 6 membered
ring having one B atom and two O atoms with the remainder carbon
atoms, and the ring may be optionally substituted with alkyl
groups, using, for example, Pd(PPh.sub.3).sub.4 or Pd(dppf)Cl.sub.2
in the presence of Na.sub.2CO.sub.3 followed by removal of the
benzoate protecting group (e.g., LiOH in an aqueous/organic solvent
mixture at 0.degree. C. to 50.degree. C.) gives compound 24.
Removal of the amine protecting group (eg. for a Boc group,
HCl/dioxane or TFA) and subsequent acylation (eg. HBTU, Hunig's
base) gives 25.
[0194] Alternatively, optional reduction of the olefin 24 (e.g.,
H.sub.2--Pd/C), followed by the removal of the amine protecting
group (e.g., for a Boc group, HCl/dioxane or TFA) and subsequent
acylation (e.g., HBTU, Hunig's base) gives 27.
[0195] Optional functionalization of the hydroxyl group of
compounds 25 or 27 may provide an entry into alternate R.sup.2
groups. For example, the alcohol may be converted to a fluoro
group, wherein compounds 26 or 28 have R.sup.2 as F and R.sup.2a as
H, by treatment with, for example, DAST. Alternatively, the alcohol
of 25 or 27 may be alkylated (e.g., MeI, NaH) to give the methoxy
analog, wherein compounds 26 or 28 have R.sup.2 as OMe and R.sup.2a
as H. Alternatively, compounds 25 or 27 may be oxidized (e.g.,
Swern-like conditions) to provide the ketone, wherein compounds 26
or 28 have R.sup.2 and R.sup.2a as oxo, which in turn could be
treated with a fluorinating agent, such as DAST or Deoxo-Fluor, in
an appropriate solvent, such as dichloromethane ("DCM") or
chloroform, to give the gem-difluoride analogue, wherein compounds
26 or 28 have R.sup.2 as F and R.sup.2a as F.
##STR00048##
[0196] Scheme 7 shows an alternative way to prepare compounds 26
and 28, wherein R.sup.1, R.sup.1a, R.sup.2 and R.sup.2a are as
defined herein. In this scheme, the pyrimidine moieties are
functionalized at an earlier stage. A Pd-mediated coupling between
the halopyrimidine 29, wherein Hal is Br, Cl or I, and an
appropriately substituted boronic acid or ester 23, wherein PG is
an amine protecting group and R.sup.m is hydrogen or alkyl
optionally substituted with OH or the two R.sup.m groups together
with the atoms to which they are attached form a 5 or 6 membered
ring having one B atom and two O atoms with the remainder carbon
atoms, and the ring may be optionally substituted with alkyl
groups, using, for example, Pd(PPh.sub.3).sub.4 or Pd(dppf)Cl.sub.2
in the presence of Na.sub.2CO.sub.3, followed by the removal of the
benzoate protecting group (e.g., LiOH in an aqueous/organic solvent
mixture at 0.degree. C. to 50.degree. C.) gives compound 30.
Removal of the amine protecting group (e.g., for a Boc group,
HCl/dioxane or TFA) and subsequent acylation (e.g., HBTU, Hunig's
base) gives 26.
[0197] Alternatively, optional reduction of the olefin 30 (e.g.,
H.sub.2--Pd/C), followed by the removal of the amine protecting
group (e.g., for a Boc group, HCl/dioxane or TFA) and subsequent
acylation (e.g., HBTU, Hunig's base) gives compound 28.
##STR00049##
[0198] Scheme 8 shows a method of generating compounds 39 and 40,
wherein R.sup.1, R.sup.1a, R.sup.2, R.sup.2a, R.sup.5 and G are as
defined herein. An appropriately substituted halobenzene 34,
wherein Hal is Br, Cl or I, is treated with a strong base, such as
BuLi, t-BuLi, Mg, etc., and addition of the newly formed anion into
the ketone of compound 33, wherein Pg is an amine protecting group,
at -100.degree. C. to 50.degree. C. gives compound 35. Compound 35
is treated with a halobenzene 36, wherein Hal is Br, Cl or I, in
the presence of a Lewis acid (e.g., AlCl.sub.3 at -20.degree. C. to
100.degree. C.) and reprotection of the amine, if necessary (e.g.,
Boc.sub.2O for a Boc-group), gives compound 37. Conversion of
compound 37 to an appropriate organometallic (e.g., treatment with
Sn.sub.2Me.sub.6, Pd(PPh.sub.3).sub.4; bispinacol ester boronate,
Pd(dppf)Cl.sub.2; or Mg) followed by a Pd-mediated coupling with
compound 38, wherein R.sup.k is halogen or NO.sub.2, using, for
example, PdCl.sub.2(PPh.sub.3).sub.2 and aqueous Na.sub.2CO.sub.3
in dioxane at room temperature to reflux, and final removal of the
amine (e.g., HCl/dioxane for a Boc-group) and the alcohol
protecting groups (e.g., LiOH in THF/H.sub.2O at 0.degree. C. to
50.degree. C.) gives compound 39.
[0199] Alternatively, a differentially functionalized pyrimidine
moiety (e.g., compound 29, wherein Hal is Br, Cl or I) could be
used in the same Pd-coupling and deprotection steps to give
compound 40.
##STR00050##
[0200] Scheme 9 shows a method of preparing compound 43. A
palladium-mediated coupling between compound 38, wherein R.sup.k is
halogen or NO.sub.2, and compound 41 using, for example,
PdCl.sub.2(dppf) and aqueous Na.sub.2CO.sub.3 in dioxane at room
temperature to reflux, followed by saponification of the esters
(e.g., LiOH or NaOH in THF/water at 0.degree. C. to reflux) gives
compound 42. The newly formed acid is treated with an appropriately
substituted primary or secondary amine under standard coupling
conditions (e.g., HBTU/DIPEA/DMF) to give compound 43.
##STR00051##
[0201] Scheme 10 shows a means of preparing compound 45, wherein
R.sup.1, R.sup.1a, R.sup.2 and R.sup.2a are as defined herein, with
a differently substituted pyrimidine moiety. A palladium-mediated
coupling between compound 29, wherein Hal is Br, Cl or I, and
compound 41 using, for example, PdCl.sub.2(dppf) and aqueous
Na.sub.2CO.sub.3 in dioxane at room temperature to reflux, followed
by saponification of the ester (e.g., LiOH or NaOH in THF/water at
0.degree. C. to reflux) gives compound 44. Treatment of the newly
formed acid with an appropriately substituted primary or secondary
amine under standard coupling conditions (e.g., HBTU/DIPEA/DMF)
gives compound 45.
##STR00052##
[0202] Scheme 11 shows a means of generating compounds 48 and 50,
wherein R.sup.1, R.sup.1a, R.sup.2 and R.sup.2a are as defined
herein. An appropriately mono-substituted diazepine 46, wherein PG
is an amine protecting group, is treated with compound 29, wherein
Hal is Br, Cl or I, and a tertiary amine base (e.g., Hunig's base)
in a suitable solvent (e.g., isopropanol) at room temperature to
reflux, followed by removal of the amine protecting group, using,
for example, in the case of a Boc-group, TFA or HCl/dioxane at
0.degree. C. to room temperature to give compound 47. Treatment of
compound 47 with an appropriately substituted acid using standard
amide forming conditions (e.g., HBTU/DIPEA/DCM at 0.degree. C. to
reflux) gives compound 48.
[0203] Alternatively, an appropriately mono-substituted diazepine
46 is treated with compound 38, wherein R.sup.k is halogen or
NO.sub.2, and a tertiary amine base (e.g., Hunig's base) in a
suitable solvent (e.g., isopropanol) at room temperature to reflux,
followed by removal of the amine protecting group, using, for
example, in the case of a Boc-group, TFA or HCl/dioxane at
0.degree. C. to room temperature to give compound 49. Compound 49
is treated with an appropriately substituted acid using standard
amide forming conditions (e.g., HBTU/DIPEA/DCM at 0.degree. C. to
reflux) followed by saponification of the ester (e.g., LiOH or NaOH
in THF/water at 0.degree. C. to reflux) gives compound 50.
##STR00053##
[0204] Scheme 12 shows a method for the formation of compounds 53
and 54, wherein R.sup.1, R.sup.1a, R.sup.2, R.sup.2a, R.sup.3,
R.sup.4, R.sup.5, R.sup.6, G, a and b are as defined herein.
Compound 51, wherein PG is an amine protecting group, is reduced
using, for example, NaBH.sub.4 in EtOH at 0.degree. C. to
50.degree. C. The resulting alcohol is then alkylated with an
appropriate amine-containing side chain (e.g.,
N(CH.sub.3).sub.2CH.sub.2CH.sub.2Cl) using, for example, a base
such as NaH in DMF at room temperature to 100.degree. C. This is
followed by deprotection of the piperidine amine using, for
example, in the case of Boc, HCl/dioxane or TFA at 0.degree. C. to
room temperature, gives compound 52. Compound 52 may then be
treated with the halopyrimidine 29, wherein Hal is Br, Cl or I, and
a tertiary amine base (e.g., Hunig's base) in, for example, DMF at
room temperature to 140.degree. C. to give compound 53.
[0205] Alternatively, compound 52 may be treated with compound 38,
wherein Hal is Br, Cl or I and R.sup.k is halogen or NO.sub.2,
under similar conditions, followed by saponification of the benzoyl
group using, for example, LiOH in THF/water at 0.degree. C. to
reflux to give compound 54.
##STR00054##
[0206] Scheme 13 describes a route to prepare compound 61, wherein
G and a are as defined herein. The amino acid 55 is reduced using,
for example, LiBH.sub.4 and ClSi(CH.sub.3).sub.3 in THF at
0.degree. C. to room temperature, followed by protection of the
amine, for example, using Boc.sub.2O, if Pg is Boc, to give
compound 56, wherein PG is an amine protecting group. Activation of
the alcohol 56 using, for example, methanesulphonyl chloride and
triethylamine in DCM at -20.degree. C. to room temperature,
followed by displacement with a protected amine, such as azide
(using, for example, sodium azide in DMF at room temperature to
120.degree. C.) and deprotection of the amine (for example, using
HCl/dioxane or TFA for a Boc group) can give the aminoazide 57.
Compound 60 may be prepared by a palladium mediated coupling
between the organozinc 59 and the chloropyrimidine 58 using, for
example, Pd(PPh.sub.3).sub.4 in THF at room temperature to reflux,
followed by saponification of the ester to give the acid 60.
Coupling of the acid 60 and the amine 57 under standard conditions
(e.g., HBTU/Hunig's base) and reduction of the azide (e.g.,
H.sub.2--Pd/C or PPh.sub.3) can give compound 61.
[0207] Alternatively, instead of the chloropyrimidine 58, an
alternatively substituted pyrimidine 29, wherein Hal is Br, Cl or I
and R.sup.1, R.sup.1a, R.sup.2 and R.sup.2a are as defined herein,
may be used using similar procedures to above, followed by
saponification, to give compound 62. A similar coupling with the
amine 57 and azide reduction then gives compound 63.
##STR00055##
[0208] Scheme 14 describes a method for the formation of compounds
67 and 68, wherein G and a are as defined herein. In this example,
compounds 58 and 59 are coupled using Pd-mediated conditions as
described in Scheme 13. Following this, the pyrimidine-1-oxide is
formed, using, for example, m-CPBA or Oxone, followed by acylation
and rearrangement with acetic anhydride at higher temperature
(e.g., 50.degree. C. to reflux) and subsequent saponification of
the acetate and ester using, for example, LiOH in THF/water at
0.degree. C. to 50.degree. C., to give the acid 64. Protection of
the acid (e.g., MeOH, trimethylsilyldiazomethane at -20.degree. C.
to room temperature) as the methyl ester, followed by acylation of
the alcohol (e.g., p-nitrobenzoyl chloride, NEt.sub.3 in DCM at
-20.degree. C. to reflux) to facilitate separation, separation
(e.g. chromatography or recrystallization) and saponification of
the benzoate and methyl esters (using, for example, LiOH in
THF/water at 0.degree. C. to 50.degree. C.) gives both compounds 65
and 66. These may be coupled with the aminoazide 57 using standard
amide forming conditions (e.g., HBTU, Hunig's base, DCM at
-20.degree. C. to reflux), followed by reduction of the azide
(e.g., H.sub.2--Pd/C or PPh.sub.3) to give compounds 67 and 68.
[0209] Optionally substituted forms of compounds 67 and 68 may be
prepared by replacing the chloropyrimidine 58 in Scheme 14, with
the optionally substituted halopyrimidine 11, or by performing the
transformations described in Scheme 17.
##STR00056##
[0210] Scheme 15 describes the preparation of compounds 73, wherein
R.sup.1, R.sup.1a, R.sup.2, R.sup.2a, R.sup.5, R.sup.6, G and a are
as defined herein. A Pd-mediated reaction between the
halopyrimidine 29, wherein Hal is Br, Cl or I, and the boronic acid
69 using, for example, PdCl.sub.2(PPh.sub.3).sub.2 and aqueous
Na.sub.2CO.sub.3 in isopropanol at room temperature to reflux,
gives the phenol 70. Compound 72 is prepared by the amine-mediated
opening of epoxide 71, followed by amine protection (e.g.,
Boc.sub.2O) if the resulting amine is primary or secondary.
Coupling of compound 72 to compound 70 under, for example,
Mitsunobu conditions (e.g., diethylazodicarboxylate and PPh.sub.3
at -40.degree. to 5.degree. C., followed by warming to temperatures
up to 50.degree. C.) and optional deprotection, if required (e.g.,
HCl/dioxane or TFA for a Boc-group), gives compound 73.
##STR00057##
[0211] Scheme 16 describes the preparation of compounds 77, wherein
R.sup.1, R.sup.1a, R.sup.2, R.sup.2a, R.sup.5, R.sup.6 and G are as
defined herein. The aniline 74 is treated with TMS-CN and an
aldehyde in the presence of an acid, such as sulphamic acid, in a
protic solvent, such as MeOH, at 0.degree. C. to 50.degree. C.,
followed by reduction of the resulting nitrile, using, for example,
LiAlH.sub.4 at -78.degree. C. to room temperature in THF, gives
compound 75. Alkylation of the amine (e.g., alkyl halide and base,
such as NaH) or reductive amination using a suitable aldehyde or
ketone in the presence of a reducing agent, such as sodium
triacetoxyborohydride, at 0.degree. C. to 50.degree. C. gives
compound 76. Compound 76 may then be converted to an appropriate
organometallic reagent by treatment with, for example, a borane,
PdCl.sub.2(dppf) and KOAc in DMSO, Sn.sub.2(CH.sub.3).sub.6 and
Pd(PPh.sub.3).sub.4 or alternatively an activated form of Mg or Zn.
This organometallic reagent may then be coupled to the
halopyrimidine 29, whrein Hal is Br, Cl or I, using a
palladium-mediated reaction (e.g., Pd(PPh.sub.3).sub.4 and aqueous
Na.sub.2CO.sub.3 in, for example, isopropanol at room temperature
to reflux) gives compound 77.
##STR00058##
[0212] Scheme 17 shows a general method for the functionalization
of the hydroxyl group of compound 79, wherein A, L, R.sup.1 and
R.sup.1a are as defined herein, providing alternative R.sup.2 and
R.sup.2a groups. The alcohol 79 may be converted to a fluoro-group
78 by treatment with a fluorinating agent such as, for example,
DAST. Alternatively, the alcohol 79 may be alkylated (e.g., a
methylating agent such as MeI and a strong base, such as NaI) to
give the methoxy analog 80 (in this instance, R.sup.2a is
hydrogen). Alternatively, compound 79 may be oxidized (e.g.,
Swern-like conditions) to provide the ketone 82, which in turn
could be treated with a fluorinating agent, such as DAST or
Deoxo-Fluor, in an appropriate solvent, such as DCM or chloroform,
to give the gem-difluoro analogue 81. Ketone 82 could also be
treated with an appropriate organometallic nucleophile, such as
MeMgBr or MeLi, to generate the tertiary alcohol 83. This may be
further fluorinated or methylated as described above to give
compounds 84 and 80 (in this instance, R.sup.2a is methyl),
respectively. Scheme 17 will generally apply to compounds of
Formula I and intermediates thereof, wherein R.sup.2 is OH and
R.sup.2a is hydrogen.
##STR00059##
[0213] Scheme 18 describes a process for preparing compound 88,
wherein R.sup.1, R.sup.1a, R.sup.2, R.sup.2a, R.sup.3, R.sup.4,
R.sup.5, R.sup.6, G, Z, a, and b are as defined herein. The acid 85
is converted to the hydrazide by treatment with, for example,
hydrazine under standard amide coupling conditions (e.g.,
1,1-carbonyldiimidazole), followed by condensation with compound
86, wherein PG is an amine protecting group, at room temperature to
150.degree. C. and removal of the protecting group (e.g., if Pg is
Boc, the use of HCl/dioxane or TFA) to give compound 87. Compound
87 is treated with compound 29, wherein Hal is Br, Cl or I, in the
presence of a base (e.g., NEt.sub.3, Hunig's base, etc.) at room
temperature to 200.degree. C., optionally in a sealed container
with or without microwave assistance, can give compound 88.
##STR00060## ##STR00061##
[0214] Scheme 19 shows a general scheme for the synthesis of
compounds 94 and 95, wherein R.sup.1, R.sup.1a, R.sup.2, R.sup.2a,
R.sup.5, R.sup.6, G and a are as defined herein. A Michael reaction
between compounds 89 and 90, wherein Hal is Br, Cl or I, in the
presence of an acid (e.g., a protic acid such as catalytic,
concentrated H.sub.2SO.sub.4) gives compound 91. This haloindole 91
may be converted to an organometallic (e.g., treatment with
Sn.sub.2(CH.sub.3).sub.6, Pd(PPh.sub.3).sub.4; bispinacol ester
boronate, Pd(dppf)Cl.sub.2; or activated Mg) followed by a
Pd-mediated coupling with compound 29 using, for example,
PdCl.sub.2(PPh.sub.3).sub.2 and aqueous Na.sub.2CO.sub.3 in dioxane
at room temperature to reflux for a Suzuki coupling, or
Ph(PPh.sub.3).sub.4 in toluene at room temperature to reflux for a
Stille coupling, gives compound 92. Reduction of the nitro group in
92 using, for example, Fe/AcOH or hydrogenation under a PtO.sub.2
catalyst gives the primary amine 93. Optional functionalization of
this amine (e.g., reductive amination, alkylation, etc.) followed
by separation of the two diastereomers then gives compounds 94 and
95.
##STR00062## ##STR00063##
[0215] Scheme 20 describes the preparation of compound 108, wherein
R.sup.1, R.sup.1a, R.sup.2, R.sup.2a, R.sup.5, R.sup.6 and G are as
defined herein. Acylation of 96, wherein Hal is Br, Cl or I, with
oxalyl chloride, followed by treatment of the resulting acid
chloride with a Lewis acid, such as AlC.sub.3, gives compound 97,
which may then be treated with aqueous ammonium hydroxide and an
oxidant, such as hydrogen peroxide, to afford the primary amide 98.
Hydrolysis of amide 98 with a base, for example, NaOH, yields
carboxylic acid 99 (a procedure is described in J. Med. Chem., 37,
2308-2314 (1994)). Compound 99 is coupled with
N,O-dimethylhydroxylamine under standard coupling conditions (e.g.,
HBTU, DIEA) to produce the amide 101. Compound 101 can react with
compound 102, which is a Grignard reagent, zinc or a lithium anion,
to produce the ketone 103. An olefination reaction of compound 103
(e.g., with methyltriphenylphosphonium bromide in the presence of a
base (for example, NaH, KOBut or NaN(Si(CH.sub.3).sub.3).sub.2))
leads to the olefin 104, which is reacted with allylamine in an
organic solvent, for example DMF, to give compound 105. Removal of
the allyl group, for example, by treatment with
1,3-dimethylpyrimidine-2,46(1H,3H,5H)-trione in the presence of
Pd(PPh.sub.3).sub.4, followed by reprotection of the free amine
with an appropriate amine protecting group, for example, Boc, using
Boc.sub.2O, gives compound 106, wherein PG is an amine protecting
group. Compound 106 may then be converted to an appropriate
organometallic reagent by treatment with, for example, a borane,
PdCl.sub.2(dppf) and KOAc in DMF, Sn.sub.2(CH.sub.3).sub.6 and
Pd(PPh.sub.3).sub.4 or alternatively an activated form of Mg or Zn.
This organometallic reagent may then be coupled to the
halopyrimidine 29, wherein Hal is Br, Cl or I, using a
palladium-mediated reaction (e.g., PdCl.sub.2(dppf) and aqueous
Na.sub.2CO.sub.3 in, for example, DMF at room temperature to
reflux) to give compound 107. Removal of protecting groups followed
by final functionalization of the unprotected free amine (e.g.,
alkylation or reductive amination to introduce new substituents)
gives rise to the final compound 108. These analogues may then be
subject to separation techniques to give the single
diastereomers.
[0216] In preparing compounds of Formula I, protection of remote
functionalities (e.g., primary or secondary amines, etc.) of
intermediates may be necessary. The need for such protection will
vary depending on the nature of the remote functionality and the
conditions of the preparation methods. Suitable amino-protecting
groups (NH-Pg) include acetyl, trifluoroacetyl, Boc,
benzyloxycarbonyl (CBz) and 9-fluorenylmethyleneoxycarbonyl (Fmoc).
The need for such protection is readily determined by one skilled
in the art. For a general description of protecting groups and
their use, see T. W. Greene, Protective Groups in Organic
Synthesis, John Wiley & Sons, New York, 1991.
Methods of Separation
[0217] It may be advantageous to separate reaction products from
one another and/or from starting materials. The desired products of
each step or series of steps is separated and/or purified
(hereinafter separated) to the desired degree of homogeneity by the
techniques common in the art. Typically such separations involve
multiphase extraction, crystallization from a solvent or solvent
mixture, distillation, sublimation, or chromatography.
Chromatography can involve any number of methods including, for
example: reverse-phase and normal phase; size exclusion; ion
exchange; high, medium and low pressure liquid chromatography
methods and apparatus; small scale analytical; simulated moving bed
(SMB) and preparative thin or thick layer chromatography, as well
as techniques of small scale thin layer and flash chromatography.
One skilled in the art will apply techniques most likely to achieve
the desired separation.
[0218] Diastereomeric mixtures can be separated into their
individual diastereomers on the basis of their physical chemical
differences by methods well known to those skilled in the art, such
as by chromatography and/or fractional crystallization. Enantiomers
can be separated by converting the enantiomeric mixture into a
diastereomeric mixture by reaction with an appropriate optically
active compound (e.g., chiral auxiliary such as a chiral alcohol or
Mosher's acid chloride), separating the diastereomers and
converting (e.g., hydrolyzing) the individual diastereoisomers to
the corresponding pure enantiomers. Enantiomers can also be
separated by use of a chiral HPLC column.
[0219] A single stereoisomer, e.g., an enantiomer, substantially
free of its stereoisomer may be obtained by resolution of the
racemic mixture using a method such as formation of diastereomers
using optically active resolving agents (Eliel, E. and Wilen, S.
"Stereochemistry of Organic Compounds," John Wiley & Sons,
Inc., New York, 1994; Lochmuller, C. H., (1975) J. Chromatogr.,
113(3):283-302). Racemic mixtures of chiral compounds of the
invention can be separated and isolated by any suitable method,
including: (1) formation of ionic, diastereomeric salts with chiral
compounds and separation by fractional crystallization or other
methods, (2) formation of diastereomeric compounds with chiral
derivatizing reagents, separation of the diastereomers, and
conversion to the pure stereoisomers, and (3) separation of the
substantially pure or enriched stereoisomers directly under chiral
conditions. See: "Drug Stereochemistry, Analytical Methods and
Pharmacology," Irving W. Wainer, Ed., Marcel Dekker, Inc., New York
(1993).
[0220] Under method (1), diastereomeric salts can be formed by
reaction of enantiomerically pure chiral bases such as brucine,
quinine, ephedrine, strychnine,
.alpha.-methyl-.beta.-phenylethylamine (amphetamine), and the like
with asymmetric compounds bearing acidic functionality, such as
carboxylic acid and sulfonic acid. The diastereomeric salts may be
induced to separate by fractional crystallization or ionic
chromatography. For separation of the optical isomers of amino
compounds, addition of chiral carboxylic or sulfonic acids, such as
camphorsulfonic acid, tartaric acid, mandelic acid, or lactic acid
can result in formation of the diastereomeric salts.
[0221] Alternatively, by method (2), the substrate to be resolved
is reacted with one enantiomer of a chiral compound to form a
diastereomeric pair (E. and Wilen, S. "Stereochemistry of Organic
Compounds", John Wiley & Sons, Inc., 1994, p. 322).
Diastereomeric compounds can be formed by reacting asymmetric
compounds with enantiomerically pure chiral derivatizing reagents,
such as menthyl derivatives, followed by separation of the
diastereomers and hydrolysis to yield the pure or enriched
enantiomer. A method of determining optical purity involves making
chiral esters, such as a menthyl ester, e.g., (-) menthyl
chloroformate in the presence of base, or Mosher ester,
.alpha.-methoxy-.alpha.-(trifluoromethyl)phenyl acetate (Jacob III.
J Org. Chem., (1982) 47:4165), of the racemic mixture, and
analyzing the .sup.1H NMR spectrum for the presence of the two
atropisomeric enantiomers or diastereomers. Stable diastereomers of
atropisomeric compounds can be separated and isolated by normal-
and reverse-phase chromatography following methods for separation
of atropisomeric naphthyl-isoquinolines (WO 96/15111).
[0222] By method (3), a racemic mixture of two enantiomers can be
separated by chromatography using a chiral stationary phase
("Chiral Liquid Chromatography" (1989) W. J. Lough, Ed., Chapman
and Hall, New York; Okamoto, J. of Chromatogr., (1990)
513:375-378). Enriched or purified enantiomers can be distinguished
by methods used to distinguish other chiral molecules with
asymmetric carbon atoms, such as optical rotation and circular
dichroism.
Administration and Pharamceutical Formulations
[0223] The compounds of the invention may be administered by any
convenient route appropriate to the condition to be treated.
Suitable routes include oral, parenteral (including subcutaneous,
intramuscular, intravenous, intraarterial, intradermal, intrathecal
and epidural), transdermal, rectal, nasal, topical (including
buccal and sublingual), vaginal, intraperitoneal, intrapulmonary
and intranasal.
[0224] The compounds may be administered in any convenient
administrative form, e.g. tablets, powders, capsules, solutions,
dispersions, suspensions, syrups, sprays, suppositories, gels,
emulsions, patches, etc. Such compositions may contain components
conventional in pharmaceutical preparations, e.g. diluents,
carriers, pH modifiers, sweeteners, bulking agents, and further
active agents. If parenteral administration is desired, the
compositions will be sterile and in a solution or suspension form
suitable for injection or infusion.
[0225] A typical formulation is prepared by mixing a compound of
the present invention and a carrier or excipient. Suitable carriers
and excipients are well known to those skilled in the art and are
described in detail in, e.g., Howard C. Ansel et al.,
Pharmaceutical Dosage Forms and Drug Delivery Systems, (8.sup.th
Ed. 2004); Alfonso R. Gennaro et al., Remington: The Science and
Practice of Pharmacy, (20.sup.th Ed. 2000); and Raymond C. Rowe,
Handbook of Pharmaceutical Excipients, (5.sup.th Ed. 2005). The
formulations may also include one or more buffers, stabilizing
agents, surfactants, wetting agents, lubricating agents,
emulsifiers, suspending agents, preservatives, antioxidants,
opaquing agents, glidants, processing aids, colorants, sweeteners,
perfuming agents, flavoring agents, diluents and other known
additives to provide an elegant presentation of the drug (i.e., a
compound of the present invention or pharmaceutical composition
thereof) or aid in the manufacturing of the pharmaceutical product
(i.e., medicament).
[0226] One embodiment of the present invention includes a
pharmaceutical composition comprising a compound of Formula I, or a
stereoisomer or pharmaceutically acceptable salt thereof. In a
further embodiment, the present invention provides a pharmaceutical
composition comprising a compound of Formula I, or a stereoisomer
or pharmaceutically acceptable salt thereof, together with a
pharmaceutically acceptable carrier or excipient.
Methods of Treatment with Compounds of the Invention
[0227] The invention includes methods of treating or preventing
disease or condition by administering one or more compounds of this
invention, or a stereoisomer or pharmaceutically acceptable salt
thereof. In one embodiment, a human patient is treated with a
compound of Formula I, or a stereoisomer or pharmaceutically
acceptable salt thereof, and a pharmaceutically acceptable carrier,
adjuvant, or vehicle in an amount to detectably inhibit AKT
activity.
[0228] In another embodiment of the present invention, a method of
treating a hyperproliferative disease in a mammal comprising
administering a therapeutically effective amount of the compound of
Formula I, or a stereoisomer or pharmaceutically acceptable salt
thereof, to the mammal is provided.
[0229] In another embodiment, a method of treating or preventing
cancer in a mammal in need of such treatment, wherein the method
comprises administering to said mammal a therapeutically effective
amount of a compound of Formula I, or a stereoisomer or
pharmaceutically acceptable salt thereof. The cancer is selected
from breast, ovary, cervix, prostate, testis, genitourinary tract,
esophagus, larynx, glioblastoma, neuroblastoma, stomach, skin,
keratoacanthoma, lung, epidermoid carcinoma, large cell carcinoma,
non-small cell lung carcinoma (NSCLC), small cell carcinoma, lung
adenocarcinoma, bone, colon, adenoma, pancreas, adenocarcinoma,
thyroid, follicular carcinoma, undifferentiated carcinoma,
papillary carcinoma, seminoma, melanoma, sarcoma, bladder
carcinoma, liver carcinoma and biliary passages, kidney carcinoma,
myeloid disorders, lymphoid disorders, hairy cells, buccal cavity
and pharynx (oral), lip, tongue, mouth, pharynx, small intestine,
colon-rectum, large intestine, rectum, brain and central nervous
system, Hodgkin's and leukemia. Another embodiment of the present
invention provides the use of a compound of Formula I, or a
stereoisomer or pharmaceutically acceptable salt thereof, in the
manufacture of a medicament for the treatment of cancer.
[0230] In another embodiment, a method of treating or preventing a
disease or disorder modulated by AKT, comprising administering to a
mammal in need of such treatment an effective amount of a compound
of Formula I, or a stereoisomer or pharmaceutically acceptable salt
thereof. Examples of such diseases and disorders include, but are
not limited to, (1) Cardiac: sarcoma (angiosarcoma, fibrosarcoma,
rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma,
lipoma and teratoma; (2) Lung: bronchogenic carcinoma (squamous
cell, undifferentiated small cell, undifferentiated large cell,
adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial
adenoma, sarcoma, lymphoma, chondromatous hamartoma, mesothelioma,
non-small cell lung, small cell lung; (3) Gastrointestinal:
esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma,
lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas
(ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma,
carcinoid tumors, vipoma), small bowel (adenocarcinoma, lymphoma,
carcinoid tumors, Karposi's sarcoma, leiomyoma, hemangioma, lipoma,
neurofibroma, fibroma), large bowel (adenocarcinoma, tubular
adenoma, villous adenoma, hamartoma, leiomyoma); (4) Genitourinary
tract: kidney (adenocarcinoma, Wilm's tumor [nephroblastoma],
lymphoma, leukemia), bladder and urethra (squamous cell carcinoma,
transitional cell carcinoma, adenocarcinoma), prostate
(adenocarcinoma, sarcoma), testis (seminoma, teratoma, embryonal
carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial
cell carcinoma, fibroma, fibroadenoma, adenomatoid tumors, lipoma);
(5) Liver: hepatoma (hepatocellular carcinoma), cholangiocarcinoma,
hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma;
(6) Bone: osteogenic sarcoma (osteosarcoma), fibrosarcoma,
malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma,
malignant lymphoma (reticulum cell sarcoma), multiple myeloma,
malignant giant cell tumor chordoma, osteochronfroma
(osteocartilaginous exostoses), benign chondroma, chondroblastoma,
chondromyxofibroma, osteoid osteoma and giant cell tumors; (7)
Nervous system: skull (osteoma, hemangioma, granuloma, xanthoma,
osteitis deformans), meninges (meningioma, meningiosarcoma,
gliomatosis), brain (astrocytoma, medulloblastoma, glioma,
ependymoma, germinoma [pinealoma], glioblastoma multifonn.
oligodendroglioma, schwannoma, retinoblastoma, congenital tumors),
spinal cord neurofibroma, meningioma, glioma, sarcoma); (8)
Gynecological: uterus (endometrial carcinoma), cervix (cervical
carcinoma, pre-tumor cervical dysplasia), ovaries (ovarian
carcinoma [serous cystadenocarcinoma, mucinous cystadenocarcinoma,
unclassified carcinoma], granulosa-thecal cell tumors,
Sertoli-Leydig cell tumors, dysgerminoma, malignant teratoma),
vulva (squamous cell carcinoma, intraepithelial carcinoma,
adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell
carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonal
rhabdomyosarcoma), fallopian tubes (carcinoma); (9) Hematologic:
blood (myeloid leukemia [acute and chronic], acute lymphoblastic
leukemia, chronic lymphocytic leukemia, myeloproliferative
diseases, multiple myeloma, myelodysplastic syndrome), Hodgkin's
disease, non-Hodgkin's lymphoma [malignant lymphoma]; (10) Skin:
advanced melanoma, malignant melanoma, basal cell carcinoma,
squamous cell carcinoma, Karposi's sarcoma, moles dysplastic nevi,
lipoma, angioma, dermatofibroma, keloids, psoriasis; (11) Adrenal
glands: neuroblastoma; (12) Breast: metastatic breast; breast
adenocarcinoma; (13) Colon; (14) Oral cavity; (15) Hairy cell
leukemia; (16) Head and neck; (17) and others including refractory
metastatic disease; Kaposi's sarcoma; Bannayan-Zonana syndrome; and
Cowden disease or Lhermitte-Duclos disease, among other kinds of
hyperproliferative disorders.
[0231] Compounds and methods of this invention can be also used to
treat diseases and conditions such as rheumatoid arthritis,
osteoarthritis, Crohn's disease, angiofibroma, ocular diseases
(e.g., retinal vascularisation, diabetic retinopathy, age-related
macular degeneration, macular degeneration, etc.), multiple
sclerosis, obesity, restenosis, autoimmune diseases, allergy,
asthma, endometriosis, atherosclerosis, vein graft stenosis,
peri-anastomatic prothetic graft stenosis, prostate hyperplasia,
chronic obstructive pulmonary disease, psoriasis, inhibition of
neurological damage due to tissue repair, scar tissue formation
(and can aid in wound healing), multiple sclerosis, inflammatory
bowel disease, infections, particularly bacterial, viral,
retroviral or parasitic infections (by increasing apoptosis),
pulmonary disease, neoplasm, Parkinson's disease, transplant
rejection (as an immunosupressant), septic shock, etc.
[0232] Another embodiment of the present invention provides the use
of a compound of Formula I, or a stereoisomer or pharmaceutically
acceptable salt thereof, in the manufacture of a medicament for the
treatment of hyperproliferative diseases. In a further embodiment,
the present invention provides the use of a compound of Formula I,
or a stereoisomer or pharmaceutically acceptable salt thereof, in
the manufacture of a medicament for the treatment of cancer. In a
further embodiment, the cancer is selected from carcinoma,
lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies.
More particular examples of such cancers include squamous cell
cancer (e.g., epithelial squamous cell cancer), lung cancer
including small-cell lung cancer, non-small cell lung cancer
("NSCLC"), adenocarcinoma of the lung and squamous carcinoma of the
lung, cancer of the peritoneum, hepatocellular cancer, gastric or
stomach cancer including gastrointestinal cancer, pancreatic
cancer, glioblastoma, cervical cancer, ovarian cancer, liver
cancer, bladder cancer, hepatoma, breast cancer, colon cancer,
rectal cancer, colorectal cancer, endometrial or uterine carcinoma,
salivary gland carcinoma, kidney or renal cancer, prostate cancer,
vulval cancer, thyroid cancer, hepatic carcinoma, anal carcinoma,
penile carcinoma, skin cancer, including melanoma, as well as head
and neck cancer.
Combination Therapy
[0233] The compounds of this invention and stereoisomers and
pharmaceutically acceptable salts thereof may be employed alone or
in combination with other therapeutic agents for treatment. The
compounds of the present invention can be used in combination with
one or more additional drugs, for example an anti-inflammatory
compound that works by a different mechanism of action. The second
compound of the pharmaceutical combination formulation or dosing
regimen preferably has complementary activities to the compound of
this invention such that they do not adversely affect each other.
Such molecules are suitably present in combination in amounts that
are effective for the purpose intended. The compounds may be
administered together in a unitary pharmaceutical composition or
separately and, when administered separately this may occur
simultaneously or sequentially in any order. Such sequential
administration may be close in time or remote in time.
EXAMPLES
[0234] In order to illustrate the invention, the following Examples
are included. However, it is to be understood that these Examples
do not limit the invention and are only meant to suggest a method
of practicing the invention. Persons skilled in the art will
recognize that the chemical reactions described may be readily
adapted to prepare a number of other compounds of the invention,
and alternative methods for preparing the compounds of this
invention are deemed to be within the scope of this invention. For
example, the synthesis of non-exemplified compounds according to
the invention may be successfully performed by modifications
apparent to those skilled in the art, e.g., by appropriately
protecting interfering groups, by utilizing other suitable reagents
known in the art other than those described, and/or by making
routine modifications of reaction conditions. Alternatively, other
reactions disclosed herein or known in the art will be recognized
as having applicability for preparing other compounds of the
invention.
[0235] In the Examples described below, unless otherwise indicated
all temperatures are set forth in degrees Celsius. Reagents were
purchased from commercial suppliers such as Sigma-Aldrich, Alfa
Aesar, or TCI, and were used without further purification unless
otherwise indicated.
[0236] The reactions set forth below were done generally under a
positive pressure of nitrogen or argon or with a drying tube
(unless otherwise stated) in anhydrous solvents, and the reaction
flasks were typically fitted with rubber septa for the introduction
of substrates and reagents via syringe. Glassware was oven dried
and/or heat dried.
[0237] Column chromatography was done on a Biotage system
(Manufacturer: Dyax Corporation) having a silica gel column or on a
silica SepPak cartridge (Waters) (unless otherwise stated). .sup.1H
NMR spectra were recorded on a Varian instrument operating at 400
MHz. .sup.1H-NMR spectra were obtained as CDCl.sub.3 [using
chloroform (7.25 ppm) as the reference standard]d.sub.6-DMSO [using
DMSO (2.50 ppm) as the reference standard], CH.sub.3OD [using
methanol (3.31 ppm) as the reference standard] or d.sub.6-acetone
[using acetone (2.05 ppm) as the reference standard] solutions
(reported in ppm). Alternatively, tetramethylsilane can be used as
an internal reference standard (0.00 ppm). When peak multiplicities
are reported, the following abbreviations are used: s (singlet), d
(doublet), t (triplet), q (quartet), m (multiplet), br (broadened),
dd (doublet of doublets), dt (doublet of triplets). Coupling
constants, when given, are reported in Hertz (Hz).
Example A
AKT-1 Kinase Assay
[0238] The activity of the compounds described in the present
invention may be determined by the following kinase assay, which
measures the phosphorylation of a fluorescently-labeled peptide by
full-length human recombinant active AKT-1 by fluorescent
polarization using a commercially available IMAP kit.
[0239] The assay materials are obtained from an IMAP AKT Assay Bulk
Kit, product #R8059, from Molecular Devices, Sunnyvale, Calif. The
kit materials include an IMAP Reaction Buffer (5.times.). The
diluted 1.times.IMAP Reaction Buffer contained 10 mM Tris-HCl, pH
7.2, 10 mM MgCl.sub.2, 0.1% BSA, 0.05% NaN.sub.3. DTT is routinely
added to a final concentration of 1 mM immediately prior to use.
Also included is IMAP Binding Buffer (5.times.), and IMAP Binding
Reagent. The Binding Solution is prepared as a 1:400 dilution of
IMAP Binding Reagent into 1.times.IMAP Binding Buffer.
[0240] The fluorescein-labeled AKT Substrate (Crosstide) has the
sequence (Fl)-GRPRTSSFAEG. A stock solution of 20 .mu.M is made up
in 1.times.IMAP Reaction Buffer.
[0241] The plates used include a Costar 3657 (382-well made of
polypropylene and having a white, v-bottom) that is used for
compound dilution and for preparing the compound-ATP mixture. The
assay plate is a Packard ProxyPlate.TM.-384 F.
[0242] The AKT-1 used is made from full-length, human recombinant
AKT-1 that is activated with PDK1 and MAP kinase 2.
[0243] To perform the assay, stock solutions of compounds at 10 mM
in dimethylsulfoxide ("DMSO") are prepared. The stock solutions and
the control compound are serially diluted 1:2 nine times into DMSO
(10 .mu.L of compound+10 .mu.L of DMSO) to give 50.times. dilution
series over the desired dosing range. Next, 2.1-.mu.L aliquots of
the compounds in DMSO are transferred to a Costar 3657 plate
containing 50 .mu.L of 10.4 .mu.M ATP in 1.times.IMAP Reaction
Buffer containing 1 mM DTT. After thorough mixing, 2.5 .mu.L
aliquots are transferred to a ProxyPlate.TM.-384 F plate.
[0244] The assay is initiated by the addition of 2.5-.mu.L aliquots
of a solution containing 200 nM of fluorescently-labeled peptide
substrate and 4 nM AKT-1. The plate is centrifuged for 1 minute at
1000 g and incubated for 60 minute at ambient temperature. The
reaction is then quenched by the addition of 15 .mu.L of Binding
Solution, centrifuged again and incubated for an additional 30
minutes at ambient temperature prior to reading on a Victor 1420
Multilabel HTS Counter configured to measure fluorescence
polarization.
[0245] Representative compounds were tested in the above assay and
found to have IC.sub.50 values of less than 10 .mu.M.
Example 1
##STR00064##
[0246]
(5R,7R)-4-(4-((S)-(4-chlorophenyl)(2-(dimethylamino)ethoxy)methyl)p-
iperidin-1-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-ol
[0247] Step 1: tert-Butyl
4-(4-chlorobenzoyl)piperidine-1-carboxylate (3.63 g, 11.2 mmol) was
dissolved in ethanol (100 mL) then sodium tetrahydroborate (424 mg,
112 mmol) was added to this solution. The reaction mixture was
stirred at room temperature for 3 h. LC-MS analysis of the reaction
mixture showed no more starting material. The crude reaction
mixture was concentrated and the residue was diluted with EtOAc
(100 mL) and washed with water (2.times.50 mL) and brine (50 mL).
The organic layer was dried over Na.sub.2SO.sub.4, filtered and
concentrated to yield tert-butyl 4-((4-chlorophenyl)(hydroxy)
methyl)piperidine-1-carboxylate (3.46 g, 94%), which was used in
the next step directly. m/z: 326 (MH.sup.+).
[0248] Step 2: To a solution of tert-butyl
4-((4-chlorophenyl)(hydroxyl)methyl) piperidine-1-carboxylate (1.25
g, 3.84 mmol) in DMF (50 mL) was added sodium hydride 60% w/w
dispersion on mineral oil (384 mg, 9.59 mmol). The reaction mixture
was stirred for 10 minutes at room temperature then
2-chloro-N,N-dimethylethanamine hydrochloride (608 mg, 4.22 mmol)
ws added. The reaction mixture was stirred at 70.degree. C. for 12
h. LC-MS of the reaction mixture showed no more starting material.
The reaction mixture was cooled to room temperature and was diluted
with EtOAc (50 mL) and washed with 10% LiCl in water (2.times.50
mL) followed by water (50 mL) and brine (50 mL). The organic layer
was dried over Na.sub.2SO4, filtered and concentrated to yield
tert-butyl
4-((4-chlorophenyl)(2-(dimethylamino)ethoxy)methyl)piperidine-1-carboxyla-
te that was purified by reversed phase HPLC followed by separation
on enantiomers by SFC. (253 mg and 246 mg, 33%) m/z: 397
(MH.sup.+).
[0249] Step 3: To a solution of tert-butyl
4-((4-chlorophenyl)(2-(dimethylamino)ethoxy)methyl)piperidine-1-carboxyla-
te (156 mg, 0.39 mmol) in methanol (1 mL) was added a solution of
4N HCl on dioxane (1 mL, 4.0 mmol). The reaction mixture was
stirred at room temperature for 2 h. LC-MS of the reaction mixture
showed no more starting material. The solvent was removed and the
(S)-2-((4-chlorophenyl)(piperidin-4-yl)methoxy)-N,N-dimethylethanamine
was used in the next step without further purification. m/z: 297
(MH.sup.+).
[0250] Step 4: To a solution of
(5R,7R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-yl
4-nitrobenzoate (65 mg, 0.196 mmol) in THF (1 mL) and water (1 mL)
at 0.degree. C. was added lithium hydroxide (19 mg, 0.39 mmol). The
reaction mixture was stirred at 0.degree. C. for 30 minutes then it
was warmed up to room temperature and stirred for 2 h. LC-MS
analysis of the reaction mixture showed no more starting material.
THF was removed and water was added (5 mL). The residue was
extracted with EtOAc (3.times.10 mL). The combined organic layers
were washed with NaHCO.sub.3 (10 mL), brine (10 mL), dried over
Na.sub.2SO.sub.4, filtered and concentrated. The crude product was
diluted with 1-butanol (2 mL) and crude
(S)-2-((4-chlorophenyl)(piperidin-4-yl)methoxy)-N,N-dimethylethanamine
was added followed by DIPEA (0.39 mL, 2.2 mmol). The reaction
mixture was heated to 135.degree. C. and was stirred for 4 h. LC-MS
of the reaction mixture showed no more starting material. The
solvent was evaporated and the residue was purified on silica gel
(0%-6% 2N NH.sub.3 in MeOH/DCM) gradient elution to yield
(5R,7R)-4-(4((S)-(4-chlorophenyl)(2-(dimethylamino)ethoxy)methyl)piperdin-
-1-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-ol as a
foam (78 mg, 98%). m/z: 445 (M); H NMR (DMSO d.sub.6): 9.33 (br s,
1H), 8.61 (s, 1H), 7.48 (d, 2H, J=8.2 Hz), 7.34 (d, 2H, J=8.2 Hz),
5.14-5.11 (m, 1H), 4.69-4.66 (m, 1H), 4.41-4.39 (m, 1H), 4.16-4.14
(m, 1H), 3.45-3.04 (m, 7H), 2.81-2.75 (m, 6H), 2.13-2.00 (m, 4H),
1.39-1.70 (m, 3H), 1.11 (d, 3H, J=6.8 Hz).
Example 2
##STR00065##
[0251]
(5R,7R)-4-(4-((R)-(4-chlorophenyl)(2-(dimethylamino)ethoxy)methyl)p-
iperidin-1-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-ol
[0252] To a solution of
(5R,7R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-yl
4-nitrobenzoate (65 mg, 0.196 mmol) in THF (1 mL) and water (1 mL)
at 0.degree. C. was added lithium hydroxide (19 mg, 0.39 mmol). The
reaction mixture was stirred at 0.degree. C. for 30 minutes then it
was warmed up to room temperature and stirred for 2 h. LC-MS
analysis of the reaction mixture showed no more starting material.
THF was removed and water was added (5 mL). The residue was
extracted with EtOAc (3.times.10 mL). The combined organic layers
were washed with NaHCO.sub.3 (10 mL), brine (10 mL), dried over
Na.sub.2SO.sub.4, filtered and concentrated. The crude product was
diluted with 1-butanol (2 mL) and crude
(R)-2-((4-chlorophenyl)(piperidin-4-yl)methoxy)-N,N-dimethylethanamine
was added followed by DIPEA (0.39 mL, 2.2 mmol). The reaction
mixture was heated to 135.degree. C. and was stirred for 4 h. LC-MS
of the reaction mixture showed no more starting material. The
solvent was evaporated and the residue was purified on silica gel
(0%-6% 2N NH.sub.3 in MeOH/DCM) gradient elution to yield
(5R,7R)-4-(4-((R)-(4-chlorophenyl)(2-(dimethylamino)ethoxy)methyl)piperid-
in-1-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-ol as a
white foam (78 mg, 98%). m/z: 445 (M+); .sup.1H NMR (DMSO d.sub.6)
.delta. (ppm) 9.46 (br s, 1H), 8.63 (s, 1H), 7.48 (d, 2H, J=8.0
Hz), 7.35 (d, 2H, J=8.1 Hz), 5.18-5.15 (m, 1H), 4.59-4.55 (m, 2H),
4.20-4.16 (m, 1H), 3.58-3.24 (m, 6H), 3.03-2.98 (m, 1H), 2.81-2.75
(m, 6H), 2.14-2.01 (m, 4H), 1.36-1.21 (m, 3H), 1.13 (d, 3H, J=6.9
Hz).
Example 3
##STR00066##
[0253]
N--((S)-1-amino-3-(2,4-dichlorophenyl)propan-2-yl)-5-((R)-5-methyl--
6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)thiophene-2-carboxamide
[0254] Step 1: (R)-(+)-Pulegone (76.12 g, 0.5 mmol), anhydrous
NaHCO.sub.3 (12.5 g) and anhydrous ether (500 mL) were added to a 1
L round-bottom flask. The reaction mixture was cooled with an
ice-bath under nitrogen. Bromine (25.62 mL, 0.5 mmol) was added
dropwise over 30 minutes. The mixture was filtered and carefully
added to NaOEt (21%, 412 mL, 1.11 mmol) in an ice-cooled bath. The
mixture was stirred at room temperature overnight, and then 5% HCl
(1 L) and ether (300 mL) were added. The aqueous phase was
extracted with ether (2.times.300 mL). The combined organic phase
was washed with water, dried and concentrated. The residue was
added to a warmed solution of semicarbazide hydrochloride (37.5 g)
and NaOAc (37.5 g) in water (300 mL). Then boiling ethanol (300 mL)
was added to give a clear solution. The mixture was refluxed for
2.5 hours and then stirred at room temperature overnight. The
mixture was treated with water (1 L) and ether (300 mL). The
aqueous phase was extracted with ether (2.times.300 mL). The
combined organic phase was washed with water, dried and
concentrated. The residue was purified by vacuum distillation
(73-76.degree. C. at 0.8 mm Hg) to give (2R)-ethyl
2-methyl-5-(propan-2-ylidene)cyclopentanecarboxylate (63 g, 64%).
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 4.13 (m, 2H), 3.38 (d,
J=16 Hz, 0.5H), 2.93 (m, 0.5H), 2.50-2.17 (m, 2H), 1.98 (m, 1H),
1.76 (m, 1H), 1.23 (m, 6H), 1.05 (m, 6H).
[0255] Step 2: (2R)-Ethyl
2-methyl-5-(propan-2-ylidene)cyclopentanecarboxylate (24 g, 0.122
mol) in ethyl acetate (100 mL) was cooled to -68.degree. C. with
dry ice/isopropanol. Ozonized oxygen (5-7 ft.sup.3h.sup.-1 of
O.sub.2) was bubbled through the solution for 3.5 hours. The
reaction mixture was flushed with nitrogen at room temperature
until the color disappeared. The ethyl acetate was removed under
vacuum, and the residue was dissolved in acetic acid (150 mL) and
cooled by ice water. Zinc powder (45 g) was then added. The
solution was stirred for 30 minutes and then filtered. The filtrate
was neutralized with 2N NaOH (1.3 L) and NaHCO.sub.3. The aqueous
phase was extracted with ether (3.times.200 mL). The organic phase
was combined, washed with water, dried and concentrated to afford
(2R)-ethyl 2-methyl-5-oxocyclopentanecarboxylate (20 g, 96%).
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 4.21 (m, 2H), 2.77 (d,
J=11.2 Hz, 1H), 2.60 (m, 1H), 2.50-2.10 (m, 3H), 1.42 (m, 1H), 1.33
(m, 3H), 1.23 (m, 3H).
[0256] Step 3: KOH (8.3 g, 147.9 mmol) in water (60 mL) was added
to a solution of a mixture of (2R)-ethyl
2-methyl-5-oxocyclopentanecarboxylate (20 g, 117.5 mmol) and
thiourea (9.2 g, 120.9 mmol) in ethanol (100 mL). The mixture was
refluxed for 10 hours. After cooling, the solvent was removed, and
the residue was neutralized with concentrated HCl (12 mL) at
0.degree. C. The mixture was then extracted with DCM (3.times.150
mL). The solvent was removed, and the residue was purified by
silica gel chromatography, eluting with hexane/ethyl acetate (2:1)
to give
(R)-2-mercapto-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-ol
(12 g, 56%). MS (APCI+) [M+H]+ 183.
[0257] Step 4: Raney Nickel (15 g) and NH.sub.4OH (20 mL) were
added to a suspension of
(R)-2-mercapto-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-ol
(12 g, 65.8 mmol) in distilled water (100 mL). The mixture was
refluxed for 3 hours and then filtered. The filtrate was
concentrated to afford
(R)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-ol (9.89 g,
99%). MS (APCI+) [M+H]+ 151.
[0258] Steps 5 and 6 describe an alternate synthesis of
(R)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-ol, starting
from (R)-ethyl 2-methyl-5-oxocyclopentanecarboxylate.
[0259] Step 5: Ammonium acetate (240 g, 3110 mmol) was added to a
solution of (R)-ethyl 2-methyl-5-oxocyclopentanecarboxylate (106 g,
623 mmol) in MeOH (1.2 L). The reaction mixture was stirred at room
temperature under nitrogen for 20 hours, after which it was
complete as determined by TLC and HPLC. The reaction mixture was
concentrated to remove MeOH. The resulting residue was dissolved in
DCM, washed twice with H.sub.2O, once with brine, dried
(Na.sub.2SO.sub.4), filtered, and concentrated to give (R)-ethyl
2-amino-5-methylcyclopent-1-enecarboxylate (102 g, 97% yield) as an
oil. LC/MS (APCI+) m/z 170 [M+H]+.
[0260] Step 6: A solution containing (R)-ethyl
2-amino-5-methylcyclopent-1-enecarboxylate (161.6 g, 955 mmol) and
ammonium formate (90.3 g, 1433 mmol) in formamide (303.5 mL, 7640
mmol) was heated to an internal temperature of 150.degree. C. and
stirred for 17 hours. The reaction mixture was cooled, and
transferred to a 2 L single nextracted flask. Excess formamidine
was then removed by high vacuum distillation. Once formamidine
stopped coming over, the remaining oil in the still pot was
dissolved in DCM and washed with brine (3.times.200 mL). The
combined aqueous washes were extracted with DCM. The combined
organic extracts were dried (Na.sub.2SO.sub.4), filtered, and
concentrated. The resulting oil was dissolved in minimal DCM, and
this solution was added using a separatory funnel to a stirred
solution of ether (about 5 volumes of ether vs. DCM solution),
causing some precipitate to form. This precipitate was removed by
filtration through a medium frit funnel which was rinsed with ether
and disposed. The filtrate was concentrated, the trituration from
ether repeated two more times and then dried on high vacuum line to
give (R)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-ol (93.2
g, 65.0% yield) as a pasty solid. LC/MS (APCI-) m/z 149.2.
[0261] Step 7: A mixture of
(R)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-ol (5.8 g,
38.6 mmol) in POCl.sub.3 (20 mL) was refluxed for 5 minutes. Excess
POCl.sub.3 was removed under vacuum, and the residue was dissolved
in DCM (50 mL). The mixture was then added to a saturated
NaHCO.sub.3 solution (200 mL). The aqueous phase was extracted with
DCM (3.times.100 mL), and the combined organic phases were dried
and concentrated. The resulting residue was purified by silica gel
chromatography, eluting with ethyl acetate to give
(R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidine (3.18
g, 49%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. .quadrature.8.81
(s, 1H), 3.47 (m, 1H), 3.20 (m, 1H), 3.05 (m, 1H), 2.41 (m, 1H),
1.86 (m, 3H), 1.47 (m, 3H).
[0262] Step 8: Pd(PPh.sub.3).sub.4 (10 mg, 0.09 mmol) was added to
a degassed solution of
(R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta-[d]-pyrimidine (300
mg, 1.78 mmol) and a 0.5M solution of
5-ethoxycarbonyl-2-thienylzinc bromide in THF (3.6 mL, 1.78 mmol).
The reaction mixture was stirred at 70.degree. C. for 16 hours and
then it was cooled to room temperature and diluted with diethyl
ether (10 mL). Water was added (5 mL) and a precipitate was formed
and filtered. The solid was washed with more diethyl ether and the
filtrate was concentrated. The resulting oil was purified on silica
gel (0%-75% EtOAc/hexanes) to yield (R)-ethyl
5-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)thiophene-2-carbox-
ylate as an oil (400 mg, 78%). m/z: 289 (MH+), .sup.1H NMR
(CDCl.sub.3): .delta. (ppm) 8.95 (s, 1H), 7.84-7.82 (m, 1H),
7.70-7.69 (m 1H), 4.39 (dq, 2H, J1=7.2 Hz, J2=1.2 Hz), 3.77
(quint., 1H, J=7.2 Hz), 3.23-1.14 (m, 1H), 3.001-2.95 (m, 1H),
2.42-2.31 (m, 1H), 1.98-1.92 (m, 1H), 1.40 (dt, 3H, J1=7.2 Hz,
J2=1.2 Hz), 1.29 (d, 3H, J=7.2 Hz).
[0263] Step 9: A 1N solution of sodium hydroxide in water (2.4 mL)
was added to a solution of (R)-ethyl
5-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)thiophene-2-carbox-
ylate (236 mg, 0.818 mmol) in ethanol (2.5 mL). The reaction
mixture was stirred at room temperature for 16 hours. LC-MS
analysis of the reaction mixture showed no more starting material.
Water (5 mL) was added, and the solution was acidified with 1N HCl
(1 mL) and extracted with EtOAc (3.times.5 mL). The combined
organic layers were washed with brine, dried over Na.sub.2SO.sub.4,
filtered and concentrated. The resulting
(R)-5-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)thiophene-2-ca-
rboxylic acid was used without further purification. m/z: 261
(MH+).
[0264] Step 10: A solution of lithium tetrahydroborate (190 mg,
8.50 mmol) in THF (4 mL) was cooled to 0.degree. C.
Chlorotrimethylsilane (2.2 mL, 17 mmol) was added dropwise. The
mixture was stirred for 20 minutes at room temperature and then
returned to 0.degree. C.
(S)-2-amino-3-(2,4-dichlorophenyl)propanoic acid (1.00 g, 4.27
mmol) was then added. The reaction was allowed to warm up slowly to
room temperature while stirring overnight. The mixture was cooled
to 0.degree. C. and quenched by slow addition of methanol (1 mL)
followed by an aqueous solution of 2N sodium hydroxide (4.2 mL,
8.54 mmol). Volatiles were removed under reduces pressure. The
slurry was diluted with water (5 mL) and extracted with DCM
(3.times.15 mL). The combined organic layers were washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated to
afford (S)-2-amino-3-(2,4-dichlorophenyl)propan-1-ol as a solid
that was used in the next step without further purification. MS m/z
220 (M+).
[0265] Step 11: di-tert-Butyldicarbonate (850 mg, 3.90 mmol) was
added to a solution of
(S)-2-amino-3-(2,4-dichlorophenyl)propan-1-ol (859 mg, 3.90 mmol)
in CHCl.sub.3 (10 mL). The reaction mixture was stirred at room
temperature for 18 hours. LC-MS analysis of the reaction mixture
showed no more starting material. The solvent was removed under
reduced pressure, and the resulting residue was purified on silica
gel (49:49:2 DCM: EtOAc: MeOH) to yield (S)-tert-butyl
1-(2,4-dichlorophenyl)-3-hydroxypropan-2-ylcarbamate as a solid
(1.17 g, 94%). MS m/z 320 (M+); .sup.1H NMR (CDCl.sub.3): .delta.
(ppm) 7.38 (s, 1H), 7.23-7.17 (m, 2H), 4.84-4.82 (m, 1H), 3.95-3.87
(m, 1H), 3.74-3.68 (m, 1H), 3.63-3.55 (m, 1H), 3.02-2.98 (m, 2H),
2.31 (br s, 1H), 1.39 (s, 9H).
[0266] Step 12: A solution of (S)-tert-butyl
1-(2,4-dichlorophenyl)-3-hydroxypropan-2-ylcarbamate (50 mg, 0.156
mmol) in DCM (1 mL) was cooled to 0.degree. C. Triethylamine (24
.mu.L, 0.172 mmol) was added, followed by methanesulfonyl chloride
(24 .mu.L, 0.312 mmol). The reaction mixture was allowed to warm up
slowly to room temperature and was stirred for 4 hours. TLC
analysis of the reaction mixture showed no more starting material.
Diethyl ether was added, and the precipitate was filtered. The
filtrate was concentrated, and the resulting residue was diluted
with DMF (0.5 mL), and sodium azide (51 mg, 0.781 mmol) was added.
The reaction mixture was heated to 100.degree. C. for 16 hours.
LC-MS of mixture showed no more starting material. Water was added
(5 mL), and the reaction mixture was extracted with diethyl ether
(3.times.5 mL). The combined organic layers were washed with brine,
dried over Na.sub.2SO.sub.4, filtered and concentrated. The crude
product was purified on silica gel (0%-70% EtOAc/hexanes gradient
elution to yield (S)-tert-butyl
1-azido-3-(2,4-dichlorophenyl)propan-2-ylcarbamate as an oil (32
mg, 59%). m/z 345 (M+); .sup.1H NMR (CDCl.sub.3) .delta. (ppm) 7.39
(s, 1H), 7.21-7.17 (m, 2H), 4.71-4.68 (m, 1H), 4.07-4.02 (m, 1H),
3.52-3.38 (m, 2H), 2.95-2.86 (m, 2H), 1.38 (s, 9H).
[0267] Step 13: TFA (0.56 mL, 7.24 mmol) was added to a solution of
(S)-tert-butyl 1-azido-3-(2,4-dichlorophenyl)propan-2-ylcarbamate
(100 mg, 0.290 mmol) in DCM (2 mL). The reaction mixture was
stirred at room temperature for 1 hour. LC-MS of the reaction
mixture showed no more starting material. The solvent was removed,
and the residue was co-evaporated with toluene (3.times.10 mL). The
(S)-1-azido-3-(2,4-dichlorophenyl)propan-2-amine was carried to the
next step without further purification. m/z 246 (MH+).
[0268] Step 14: (S)-1-Azido-3-(2,4-dichlorophenyl)propan-2-amine
(33 mg, 0.134 mmol) was added to a solution of
(R)-5-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)thiophene-2-ca-
rboxylic acid (35 mg, 0.134 mmol) in DCM (0.7 mL). HBTU (56 mg,
0.148 mmol) was added, followed by DIPEA (0.23 mL, 1.34 mmol). The
reaction mixture was stirred at room temperature for 1 hour. LC-MS
of the reaction mixture showed no more starting material. The
solvent was removed, and the resulting residue was purified on
silica gel (0%-50% EtOAc/hexanes) gradient elution to yield
N--((S)-1-azido-3-(2,4-dichlorophenyl)propan-2-yl)-5-((R)-5-methyl-6,7-di-
hydro-5H-cyclopenta[d]pyrimidin-4-yl)thiophene-2-carboxamide as an
oil (59 mg, 90%). m/z 486 (MH+).
[0269] Step 15: 10% Pd/C (6 mg) was added to a solution of
N--((S)-1-azido-3-(2,4-dichlorophenyl)propan-2-yl)-5-((R)-5-methyl-6,7-di-
hydro-5H-cyclopenta[d]pyrimidin-4-yl)thiophene-2-carboxamide (59
mg, 0.121 mmol) in methanol (2.5 mL). The solution was put under
vacuum and purged with H.sub.2 (3.times.), and then the reaction
mixture was stirred under a hydrogen atmosphere for 2 hours. LC-MS
analysis of the reaction mixture showed no more starting material.
The reaction mixture was filtered and concentrated to yield
N--((S)-1-amino-3-(2,4-dichlorophenyl)propan-2-yl)-5-((R)-5-methyl-6,7-di-
hydro-5H-cyclopenta[d]pyrimidin-4-yl)thiophene-2-carboxamide as a
foam (53 mg, 95%). m/z 461 (M+); .sup.1H NMR (DMSO d.sub.6) .delta.
(ppm) 8.91 (s, 1H), 8.58-8.56 (m, 1H), 7.90 (br s, 2H), 7.84 (d,
1H, J=4.3 Hz), 7.78 (d, 1H, J=4.1 Hz), 7.60 (m, 1H), 7.39-7.33 (m,
2H), 4.55-4.42 (m, 1H), 3.87-3.77 (m, 1H), 3.19-2.84 (m, 6H),
2.33-2.26 (m, 1H), 1.88-1.82 (m, 1H), 1.18 (d, 3H, J=6.9 Hz).
Example 4
##STR00067##
[0270]
N--((S)-1-amino-3-(2,4-dichlorophenyl)propan-2-yl)-5-((5R,7R)-7-hyd-
roxy-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)thiophene-2-carbo-
xamide
[0271] Step 1: m-CPBA (1.06 g, 6.12 mmol) was added portionwise to
a solution of (R)-methyl
5-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)thiophene-2-carbox-
ylate (883 mg, 3.06 mmol) in CHCl.sub.3 (20 mL) at 0.degree. C. The
reaction mixture was stirred at 0.degree. C. for 10 minutes then it
was warmed up to room temperature and stirred for 18 hours. LC-MS
analysis of the reaction mixture showed no more starting material.
The reaction mixture was cooled to 0.degree. C., and a solution of
Na.sub.2S.sub.2O.sub.3 (968 mg, 6.12 mmol) in water (5 mL) was
added dropwise, followed by a solution of Na.sub.2CO.sub.3 (649 mg,
6.12 mmol) in water (10 mL). The reaction mixture was stirred for
30 minutes at 0.degree. C., and then it was warmed up to room
temperature and extracted with CHCl.sub.3 (3.times.25 mL). The
combined organic layers were washed with brine, dried over
Na.sub.2SO.sub.4, filtered and concentrated to give the N-oxide as
an oil. The crude product was dissolved in acetic anhydride (5.8
mL, 61.2 mmol), and the solution was heated to 90.degree. C. for 2
hours. The excess acetic anhydride was then removed under reduced
pressure, and the residue was dissolved in DCM (20 mL) and poured
slowly into a stirred aqueous saturated solution of
Na.sub.2CO.sub.3 cooled to 0.degree. C. The mixture was extracted
with DCM (3.times.10 mL). The combined organic layers were washed
with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated
to yield an oil. The oil was dissolved in THF (15 mL), and a
solution of LiOH (366 mg, 7.65 mmol) in water (2.2 mL) was added.
The reaction mixture was stirred at room temperature under N.sub.2
for 16 hours. LC-MS analysis of the reaction mixture showed no more
starting material. Water was added (10 mL), and the reaction
mixture was extracted with EtOAc (3.times.10 mL). The combined
organic layers were washed with brine, dried over Na.sub.2SO.sub.4,
filtered and concentrated to yield a solid that was purified by
reversed phase HPLC to give
(R)-5-(7-hydroxy-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)thio-
phene-2-carboxylic acid (336 mg, 40%) as a mixture of
diastereoisomers. m/z 277 (MH.sup.+).
[0272] Step 2: Trimethylsilyldiazomethane (1.10 mmol) was added
dropwise to a suspension of
(R)-5-(7-hydroxy-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)thio-
phene-2-carboxylic acid (200 mg, 0.723 mmol) in MeOH (5 mL) and
Et.sub.2O (5 mL). The reaction mixture was stirred for 1 hour at
room temperature, and then the solvent was removed under reduced
pressure. The residue was diluted with DCM (10 mL) and then cooled
to 0.degree. C. Triethylamine (131 .mu.L, 0.94 mmol) was added,
followed by the addition of p-nitrobenzoyl chloride (148 mg, 0.792
mmol). The reaction mixture was allowed to warm up slowly to room
temperature and was stirred for 4 hours. TLC analysis of the
reaction mixture showed no more starting material. The reaction
mixture was quenched by the addition of saturated aqueous solution
of NaHCO.sub.3 (5 mL), and the reaction mixture was extracted with
DCM (3.times.10 mL). The combined organic layers were washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated. The
crude product was purified on silica gel (10%-45% EtOAc/hexanes)
gradient elution to yield
5-((5R,7R)-5-methyl-7-(4-nitrobenzoyloxy)-6,7-dihydro-5H-cyclopenta[d]pyr-
imidin-4-yl)thiophene-2-carboxylate as a solid (32 mg, 10%). m/z
440 (MH.sup.+); .sup.1H NMR (CDCl.sub.3) .delta. (ppm) 9.12 (s,
1H), 8.31-8.26 (m, 4H), 7.87 (d, 1H, J=4.0 Hz), 7.76 (d, 1H, J=4.0
Hz), 6.63 (dd, 1H, J.sub.1=7.7 Hz, J.sub.2=7.8 Hz), 3.95 (s, 3H),
2.71-2.65 (m, 1H), 2.51-2.43 (m, 1H), 1.43 (d, 3H, J=7.1 Hz).
[0273] Step 3: Lithium hydroxide (6 mg, 0.146 mmol) was added to a
solution of
5-((5R,7R)-5-methyl-7-(4-nitrobenzoyloxy)-6,7-dihydro-5H-cyclopenta[d]pyr-
imidin-4-yl)thiophene-2-carboxylate (32 mg, 0.073 mmol) in THF (0.5
mL) and water (0.5 mL). The reaction mixture was stirred at room
temperature for 1 hour. LC-MS of the reaction mixture showed no
more starting material. THF was removed under reduced pressure, and
1N HCl (5 mL) was added. The reaction mixture was extracted with
EtOAc (3.times.5 mL). The combined organic layers were washed with
brine, dried over Na.sub.2SO.sub.4, filtered and concentrated to
give
5-((5R,7R)-7-hydroxy-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)-
thiophene-2-carboxylic acid. The crude mixture was used directly in
the next step without further purification.
[0274] Step 4: (S)-1-Azido-3-(2,4-dichlorophenyl)propan-2-amine (20
mg, 0.0724 mmol) was added to a solution of
5-((5R,7R)-7-hydroxy-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)-
thiophene-2-carboxylic acid (35 mg, 0.145 mmol) in DCM (0.5 mL).
DMF (125 .mu.L) was added to solubilize the starting material
completely. DIPEA (126 .mu.L, 0.724 mmol) was added, followed by
the addition of HBTU (30 mg, 0.079 mmol). The reaction mixture was
stirred at room temperature for 1 hour. LC-MS of the reaction
mixture showed no more starting material. The solvent was removed,
and the resulting residue was purified by reversed phase HPLC to
yield
N--((S)-1-azido-3-(2,4-dichlorophenyl)propan-2-yl)-5-((5R,7R)-7-hydroxy-5-
-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)thiophene-2-carboxamide
product as a solid (4 mg, 10%). m/z 504 (MH.sup.+).
[0275] Step 5: 10% w/w Pd/C (1 mg) was added to a solution of
N--((S)-1-azido-3-(2,4-dichlorophenyl)propan-2-yl)-5-((5R,7R)-7-hydroxy-5-
-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)thiophene-2-carboxamide
(4.0 mg, 0.008 mmol) in MeOH (0.5 mL). The solution was put under
vacuum and purged with H.sub.2 (3.times.). The reaction mixture was
then stirred under a hydrogen atmosphere for 2 hours. LC-MS
analysis of the reaction mixture showed no more starting material.
The reaction mixture was filtered and then concentrated to yield
N--((S)-1-amino-3-(2,4-dichlorophenyl)propan-2-yl)-5-((5R,7R)-7-hydroxy-5-
-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)thiophene-2-carboxamide
as a solid (3.7 mg, 98%). m/z 477 (M+). .sup.1H NMR (DMSO d.sub.6)
.delta. (ppm) 8.91 (s, 1H), 8.58-8.56 (m, 1H), 7.90 (br s, 2H),
7.84 (d, 1H, J=4.3 Hz), 7.78 (d, 1H, J=4.1 Hz), 7.60 (m, 1H),
7.39-7.33 (m, 2H), 5.22-5.17 (m, 1H), 4.59-4.50 (m, 1H), 3.87-3.77
(m, 1H), 3.19-2.84 (m, 5H), 2.33-2.26 (m, 1H), 1.88-1.82 (m, 1H),
1.13 (d, 3H, J=6.9 Hz).
Example 5
##STR00068##
[0276]
(5R,7R)-4-(4-(4-(4-chlorophenyl)piperidin-4-yl)phenyl)-5-methyl-6,7-
-dihydro-5H-cyclopenta[d]pyrimidin-7-ol
[0277] Step 1: Ammonium acetate (240 g, 3110 mmol) was added to a
solution of (R)-ethyl 2-methyl-5-oxocyclopentanecarboxylate (106.0
g, 622.8 mmol) in MeOH (1.2 L). The reaction mixture was stirred at
room temperature under nitrogen for 20 hours. The reaction was
complete as determined by TLC and HPLC. The reaction mixture was
concentrated to remove MeOH. The resulting residue was dissolved in
DCM, washed twice with H.sub.2O, once with brine, dried
(Na.sub.2SO.sub.4), filtered, and concentrated to give (R)-ethyl
2-amino-5-methylcyclopent-1-enecarboxylate (102 g, 97% yield) as an
oil. LC/MS (APCI+) m/z 170 [M+H]+.
[0278] Step 2: A solution containing (R)-ethyl
2-amino-5-methylcyclopent-1-enecarboxylate (161.6 g, 955.0 mmol)
and ammonium formate (90.3 g, 1430 mmol) in formamide (303.5 mL,
7640 mmol) was heated to an internal temperature of 150.degree. C.
and stirred for 17 hours. The reaction mixture was cooled, and
transferred to a 2 L single nextracted flask. Then excess
formamidine was removed by high vacuum distillation. Once
formamidine stopped coming over, the remaining oil in the still pot
was dissolved in DCM and washed with brine (3.times.200 mL). The
combined aqueous washes were extracted with DCM. The combined
organic extracts were dried (Na.sub.2SO.sub.4), filtered, and
concentrated. The resulting oil was dissolved in minimal DCM, and
this solution was added using a separatory funnel to a stirred
solution of ether (about 5 volumes of ether vs. DCM solution),
causing some precipitate to form. This precipitate was removed by
filtration through a medium frit funnel, which was rinsed with
ether and disposed. The filtrate was concentrated, the trituration
from ether repeated two more times and then dried on high vacuum
line to give
(R)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-ol (93.23 g,
65.00% yield) as a pasty solid. LC/MS (APCI-) m/z 149.2.
[0279] Step 3: Neat POCl.sub.3 (463.9 mL, 5067 mmol) was added
slowly by addition funnel to a 0.degree. C. solution of
(R)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-ol (152.2 g,
1013 mmol) in DCE (1.2 L). After the addition was complete, the
reaction mixture was warmed to room temperature. The reaction
mixture was then heated to reflux and stirred for 70 minutes. The
reaction was complete as determined by HPLC. The reaction mixture
was cooled to room temperature, and the excess POCl.sub.3 was
quenched in 4 portions as follows: The reaction mixture was
transferred to a separatory funnel and dripped into a beaker
containing ice and a saturated NaHCO.sub.3 solution cooled in an
ice bath. Once the addition of each portion of the reaction mixture
was completed, the quenched mixture was stirred 30 minutes to
ensure complete destruction of POCl.sub.3 prior to transfer to
separatory funnel. The mixture was transferred to the separatory
funnel and extracted twice with DCM. The combined extracts were
dried (Na.sub.2SO.sub.4), filtered, and concentrated. The crude was
purified on silica gel as follows: silica gel (1 kg) was slurried
in 9:1 hexane:ethyl acetate onto a 3 L fritted funnel, the silica
settled under vacuum and topped with sand. The crude was loaded
with a DCM/hexane mixture, and the compound was eluted using 1 L
sidearm flasks under vacuum to give
(R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidine (104.4
g, 61.09% yield) as an oil.
[0280] Step 4: Solid 77% maximum m-CPBA (12 g, 53 mmol) was added
portionwise to a 0.degree. C. solution of
(R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidine (5 g,
30 mmol) in CHCl.sub.3 (80 mL). The reaction mixture was stirred at
room temperature overnight. The reaction mixture was cooled to
0.degree. C., and then treated by the addition of NaHCO.sub.3 (25
g, 196 mmol) slurry in water (100 mL). This was followed by the
dropwise addition of Na.sub.2CO.sub.3 (14 g, 128 mmol) in water
(100 mL). The reaction mixture was stirred for 30 minutes. The
aqueous phase was extracted with CHCl.sub.3. The organic phase was
dried with MgSO.sub.4 and concentrated under reduced pressure at
low temperature (<25.degree. C.) to afford the crude
(R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidine
1-oxide (5.5 g, 100%), which was used in the next step without
further purification.
[0281] Step 5: A solution of the
(R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidine
1-oxide (5.5 g, 29.8 mmol) in Ac.sub.2O (40.5 mL, 429 mmol) was
heated to 110.degree. C. for 3 hours. After cooling, the acetic
anhydride was evaporated, and the resulting residue was taken up in
DCM. This solution was added to a stirring cold solution of
saturated NaHCO.sub.3. The layers were extracted with DCM, dried
MgSO.sub.4, filtered, and concentrated. The crude oil was
chromatographed (Biotage) eluting with 20% EtOAc/hexane, to provide
(5R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-yl
acetate (3.0 g, 44.4%).
[0282] Step 6: A solution of
(5R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-yl
acetate (11.86 g, 52.33 mmol) in 2:1 THF:H.sub.2O (270 mL) was
cooled to 0.degree. C. and treated with lithium hydroxide hydrate
(3.95 g, 94.2 mmol). The reaction was stirred at room temperature
for 4 hours. The reaction was concentrated and diluted with water
and acidified with 6N HCl to a pH of 6. The aqueous layer was
extracted with EtOAc. The combined organics were washed with brine,
dried MgSO.sub.4, filtered, and concentrated. The crude residue was
chromatographed (Biotage 65) eluting with 30-50% EtOAc/hexane to
give
(5R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-ol
(6.09 g, 63%).
[0283] Step 7: Solid 4-nitrobenzoyl chloride (6.73 g, 36.3 mmol)
was added to a stirred solution of
(5R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-ol
(6.09 g, 33.0 mmol) and NEt.sub.3 (5.98 mL, 42.9 mmol) in DCM (165
mL) at 0.degree. C. The reaction was warmed to room temperature and
then stirred at room temperature for 3 hours. The reaction was
diluted with DCM and saturated aqueous NaHCO.sub.3. The combined
extracts were washed with brine, dried MgSO.sub.4, filtered, and
concentrated. The crude residue was chromatographed eluting with
10-14% EtOAc/hexane several times to give
(5R,7S)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-yl
4-nitrobenzoate (3.96 g, 36%) and
(5R,7R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-yl
4-nitrobenzoate (5.92 g, 54%).
[0284] Step 8: n-Butyl lithium (16 mL, 2.5M solution in hexanes,
39.9 mmol) was added to a solution of 1-bromo-4-chlorobenzne (8.62
g, 45.0 mmol) in anhydrous THF (100 mL) at -78.degree. C. under
nitrogen. After 30 minutes at -78.degree. C., 1-Boc-piperidone
(6.62 g, 33.2 mmol) was added dropwise as a solution in anhydrous
THF (5 mL). After 30 minutes at -78.degree. C., the reaction was
quenched with a saturated NH.sub.4Cl solution (100 mL), extracted
with EtOAc (2.times.100 mL). The combined organics were dried
(Na.sub.2SO.sub.4), filtered and concentrated. The crude product
was purified by silica gel chromatography, eluting with
EtOAc/hexane (0-40%) to give tert-butyl
4-(4-chlorophenyl)-4-hydroxypiperidine-1-carboxylate (7.64 g, 74%
yield). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 7.38 (d, J=7.2
Hz, 2H), 7.15 (d, J=7.3 Hz, 2H), 3.94 (br s, 2H), 3.18 (br s, 2H),
1.89 (br s, 2H), 1.46 (s, 11H). LCMS: M+1 312.2.
[0285] Step 9: Solid aluminum trichloride (0.68 mL, 5.1 mmol) was
added to a solution of tert-butyl
4-(4-chlorophenyl)-4-hydroxypiperidine-1-carboxylate (0.40 g, 1.3
mmol) in bromobenzene (5.4 mL, 51.3 mmol) at 0.degree. C. under
nitrogen. After being stirred at 0.degree. C. for 4 hours, the
mixture was quenched with ice and concentrated under reduced
pressure. The mixture was dissolved in 1N LiOH (30 mL) and
extracted with EtOAc (2.times.30 mL). The combined organics were
dried with Na.sub.2SO.sub.4, filtered and concentrated. The crude
product was directly taken up in EtOAc (4 mL) and aqueous
Na.sub.2CO.sub.3 solution (2M, 4 mL). Di-tert-butyldicarbonate
(0.851 g, 3.9 mmol) was added in one portion. The mixture was
stirred at 23.degree. C. overnight. The next day, two layers were
separated, and the aqueous layer was further extracted with EtOAc
(2.times.10 mL). The combined organics were dried over
Na.sub.2SO.sub.4, concentrated, and purified by column
chromatography (0-40% EtOAc/hexanes) to give tert-butyl
4-(4-bromophenyl)-4-(4-chlorophenyl)piperidine-1-carboxylate (0.302
g, 51% yield). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 7.46 (d,
J=7.2 Hz, 2H), 7.32 (d, J=6.9 Hz, 2H), 7.19 (d, J=7.0 Hz, 2H), 7.12
(d, J=7.3 Hz, 2H), 3.54 (br s, 2H), 2.35 (br s, 2H), 1.54 (br s,
2H), 1.43 (s, 11H). LCMS: M+1 452.2.
[0286] Step 10: Bispinacol ester boronate (93 mg, 0.37 mmol) was
added under nitrogen to a solution of tert-butyl
4-(4-bromophenyl)-4-(4-chlorophenyl)piperidine-1-carboxylate (150
mg, 0.33 mmol) in 1,4-dioxane (5 mL) at 23.degree. C. Potassium
acetate (98 mg, 1.0 mmol) was then added, and finally
1,1'-bis(diphenylphosphino)ferrocenepalladium (II) chloride (10 mg,
0.02 mmol) was added. The mixture was heated to 100.degree. C.
under nitrogen. After 3 hours, the reaction was cooled to room
temperature, diluted with H.sub.2O (10 mL), and extracted with
EtOAc (2.times.10 mL). The combined organics were dried
(Na.sub.2SO.sub.4), filtered and concentrated. The crude material
was purified by column chromatography (gradient from 100% hexane to
4:6 EtOAc:hexanes) to give the boronate (120 mg, 72% yield).
(5R,7R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-yl
4-nitrobenzoate (82 mg, 0.246 mmol) was added to this boronate (120
mg, 0.24 mmol), followed by the addition of 1,4-dioxane (3 mL) and
aqueous Na.sub.2CO.sub.3 solution (1M, 0.3 mL). The mixture was
flushed with nitrogen and bis(triphenylphosphine)palladium(II)
chloride (8.6 mg, 0.123 mmol) was added in one portion. The mixture
was heated at 100.degree. C. under nitrogen for 12 hours. Then the
reaction mixture was diluted with 0.1N LiOH solution (10 mL) and
extracted into EtOAc (2.times.10 mL). The combined organic extracts
were dried over Na.sub.2SO.sub.4, filtered, concentrated under
reduced pressure, and purified by column chromatography (0-100%
EtOAc/hexanes) to give tert-butyl
4-(4-chlorophenyl)-4-(4-((5R,7R)-5-methyl-7-(4-nitrobenzoyloxy)-6,7-dihyd-
ro-5H-cyclopenta[d]pyrimidin-4-yl)phenyl)piperidine-1-carboxylate
(36 mg, 29% yield) as an oil.
[0287] Step 11: tert-Butyl
4-(4-chlorophenyl)-4-(4-((5R,7R)-5-methyl-7-(4-nitrobenzoyloxy)-6,7-dihyd-
ro-5H-cyclopenta[d]pyrimidin-4-yl)phenyl)piperidine-1-carboxylate
(36 mg, 0.069 mmol) was dissolved in DCM (3 mL) and cooled to
0.degree. C. Then TFA (1.5 mL) was added dropwise. The resulting
solution was stirred at 0.degree. C. for 1 hour, concentrated under
reduced pressure, dissolved in DMF (1 mL) and purified by reverse
phase HPLC (0-100% AcCN/H.sub.2O) to give
(5R,7R)-4-(4-(4-(4-chlorophenyl)piperidin-4-yl)phenyl)-5-methyl-6-
,7-dihydro-5H-cyclopenta[d]pyrimidin-7-ol di-TFA salt (9 mg, 31%
yield). .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. 9.09 (s, 1H),
7.88 (d, J=8.8 Hz, 2H), 7.52 (d, J=8.4 Hz, 2H), 7.42 (q, J=4.8 Hz,
4H), 5.64 (d, J=6.0 Hz, 1H), 5.06 (q, J=6.8 Hz, 1H), 3.85 (br s,
1H), 3.05 (br s, 3H), 2.89 (m, 2H), 2.61 (br s, 3H), 2.11 (m, 2H),
1.16 (t, J=6.4 Hz, 3H), 1.04 (d, J=6.6 Hz, 3H). LCMS: M+1
420.4.
Example 6
##STR00069##
[0288]
N--((R)-2-(4-chlorophenyl)-2-(6-((R)-5-methyl-6,7-dihydro-5H-cyclop-
enta[d]pyrimidin-4-yl)-1 H-indol-3-yl)ethyl)propan-2-amine
[0289] Step 1: 1-((1E)-2-Nitrovinyl)-4-chlorobenzene (0.206 g, 1.12
mmol) was added to a solution of 6-bromoindole (0.200 g, 1.02 mmol)
in methanol (5 mL), followed by the addition of sufamic acid (39.6
mg, 0.40 mmol). The mixture was stirred at 80.degree. C. for 16
hours. The mixture was concentrated then diluted with EtOAc (15 mL)
and washed with H.sub.2O (15 mL). The organic layer was dried
(Na.sub.2SO.sub.4), filtered and concentrated. The crude product
was purified by silica gel chromatography, eluting with
EtOAc/hexane (0-50%) to give
6-bromo-3-(1-(4-chlorophenyl)-2-nitroethyl)-1H-indole (0.175 g,
45.2%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 8.12 (s, 1H),
7.51 (s, 1H), 7.28 (d, J=8.5 Hz, 2H), 7.25-7.20 (m, 3H), 7.17 (d,
J=8.5 Hz, 1H), 7.15 (s, 1H), 5.12 (t, 1H), 5.01 (q, 1H), 4.88 (q,
1H).
[0290] Step 2: A solution containing
6-bromo-3-(1-(4-chlorophenyl)-2-nitroethyl)-1H-indole (0.210 g,
0.553 mmol), bispinacol ester boronate (168 mg, 0.664 mmol) and
potassium acetate (0.163 g, 1.66 mmol) in 1,4-dioxane (5.00 mL) was
deoxygenated, and then
[1,1'-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) complex
with dichloromethane (22.6 mg, 0.0276 mmol, 1:1) was added. The
mixture was heated at 80.degree. C. for 15 hours. The mixture was
cooled to ambient temperature, diluted with H.sub.2O (10 mL) and
extracted with EtOAc (2.times.10 mL). The combined organic were
dried (Na.sub.2SO.sub.4), filtered and concentrated. The crude
product was purified by silica gel chromatography, eluting with
EtOAc/hexane (0-50%) to give pure boronate (0.146 g, 74.5%).
(R)-4-Chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidine (52.4
mg, 0.311 mmol), Bis(triphenylphosphine)palladium(II) chloride
(17.5 mg, 0.0249 mmol) were added to the boronate (0.146 g, 0.342
mmol), followed by the addition of acetonitrile (0.933 mL) and
aqueous KOAc solution (0.933 mL, 1M). The mixture was cooled to
ambient temperature, diluted with H.sub.2O (10 mL) and extracted
with EtOAc (2.times.10 mL). The combined organic were dried
(Na.sub.2SO.sub.4), filtered and concentrated. The crude product
was purified by silica gel chromatography, eluting with
EtOAc/hexane (0-100%) to give
(5R)-4-(3-(1-(4-chlorophenyl)-2-nitroethyl)-1H-indol-6-yl)-5-methyl-6,7-d-
ihydro-5H-cyclopenta[d]pyrimidine (0.115 g, 85.4%). LCMS: M+1
433.4.
[0291] Step 3: A suspension of nickel chloride hexahydrate (31.6
mg, 0.133 mmol) in methanol (10 mL, 200 mmol) was sonicated until
complete dissolution. Sodium tetrahydroborate (15.1 mg, 0.398 mmol)
was added in small portion to this stirring solution at room
temperature. As more and more NaBH.sub.4 was added, a precipitate
formed. Upon complete addition of NaBH.sub.4, the suspension was
allowed to stir at room temperature for 30 minutes. A solution of
(5R)-4-(3-(1-(4-chlorophenyl)-2-nitroethyl)-1H-indol-6-yl)-5-methyl-6,7-d-
ihydro-5H-cyclopenta[d]pyrimidine (115 mg, 0.266 mmol) in MeOH (2
mL) was added to this stirring suspension. Sodium tetrahydroborate
(35.2 mg, 0.930 mmol) was carefully added in small portions to this
stirring suspension. The reaction mixture was stirred for 1 hour at
room temperature. The reaction mixture was filtered thru celite,
and the filtrate was treated with aqueous NH.sub.4OH (10 mL, 20%
solution NH.sub.4OH). The filtrate was then extracted with
CHCl.sub.3 (4.times.10 mL). The combined organics were dried
(Na.sub.2SO.sub.4), filtered and concentrated. Crude
2-(4-chlorophenyl)-2-(6-((R)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimid-
in-4-yl)-1H-indol-3-yl)ethanamine was carried to next step. LCMS:
M+1 403.5.
[0292] Step 4: Acetone (0.0195 mL, 0.266 mmol) was added to a
solution of
2-(4-chlorophenyl)-2-(6-((R)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimid-
in-4-yl)-1H-indol-3-yl)ethanamine (107 mg, 0.266 mmol) and
N,N-diisopropylethylamine (0.139 mL, 0.797 mmol) in methylene
chloride (2.0 mL). The reaction mixture was stirred for 10 minutes,
and then sodium triacetoxyborohydride (0.0188 g, 0.0885 mmol) was
added. After stirring for 2.5 hours, saturated NaHCO.sub.3 was
added, and the mixture was stirred vigorously 10 minutes. The
mixture was extracted with DCM (3.times.10 mL). The combined
organic were dried (Na.sub.2SO.sub.4), filtered and concentrated.
The crude product was purified by SFC chiral chromotagraphy (Chiral
OJH (21.2.times.250 mm) 25% methanol+0.1% TEA) to give
N--((R)-2-(4-chlorophenyl)-2-(6-((R)-5-methyl-6,7-dihydro-5H-cyclope-
nta[d]pyrimidin-4-yl)-1H-indol-3-yl)ethyl)propan-2-amine (19.8 mg,
16.8%) as the first peak. LCMS: M+1 445.3. .sup.1H NMR (CDCl.sub.3,
400 MHz) .delta. 11.16 (s, 1H), 8.93 (s, 1H), 7.93 (s, 1H),
7.49-7.46 (m, 3H), 7.38 (d, J=8.4 Hz, 2H), 7.31 (d, J=8.5 Hz, 2H),
4.36 (t, 1H), 3.89 (m, 1H), 3.23 (m, 1H), 3.14-2.99 (m, 2H),
2.94-2.86 (m, 1H), 2.77 (m, 1H), 2.34 (m, 1H), 1.68 (m, 1H), 0.96
(d, 6H), 0.92 (d, 3H).
Example 7
##STR00070##
[0293]
(R)--N-(2-(4-chlorophenyl)-2-(4-(5-methyl-6,7-dihydro-5H-cyclopenta-
[d]pyrimidin-4-yl)phenoxy)ethyl)propan-2-amine
[0294] Step 1: 2-Propanamine (1.71 mL, 20.0 mmol) was added to a
mixture of 2-(4-chlorophenyl)oxirane (2.69 g, 16.7 mmol) and water
(6.67 mL). The mixture was stirred at room temperature. After 24
hours, additional 2-propanamine (1.13 mL, 13.3 mmol) was added.
After a total of 40 hours, water (15 mL) was added. The contents
were extracted with ether (4.times.50 mL). The combined ether
solutions were dried (Na.sub.2SO.sub.4). After filtration and
evaporation of solvents, the material was dissolved in methylene
chloride (100 mL). Di-tert-butyldicarbonate (4.01 g, 18.4 mmol) was
added, followed by triethylamine (2.56 mL, 18.4 mmol). The mixture
was stirred at room temperature for 20 hours. More Boc.sub.2O (2 g)
was added. After 4 hours, imidazole (1.136 g, 16.68 mmol) was
added. After 20 minutes, the contents were diluted with DCM (50
mL), washed with 1.0M NaH.sub.2PO.sub.4 (2.times.50 mL), and dried
(Na.sub.2SO.sub.4). The crude was purified by flash chromatography
to give tert-butyl
2-(4-chlorophenyl)-2-hydroxyethyl(isopropyl)carbamate (3.32 g,
63%).
[0295] Step 2:
(R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidine (137
mg, 0.812 mmol) and 4-hydroxybenzeneboronic acid (123 mg, 0.894
mmol) was dissolved in isopropyl alcohol (1.6 mL). Sodium carbonate
in water (0.97 mL, 1.0M) was added slowly, followed by
bis(triphenylphosphine)palladium chloride (26.5 mg, 0.0378 mmol).
The mixture was stirred at 100.degree. C. (bath) for 16 hours. The
contents were diluted with water (5 mL) and EtOAc (10 mL). The
aqueous layer was separated and extracted with EtOAc (2.times.5
mL). The combined EtOAc solutions were washed with brine (5 mL) and
dried (Na.sub.2SO.sub.4). The crude was purified by flash
chromatography to give
(R)-4-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)phenol
(160 mg, 87%) as viscous oil which solidified upon standing.
[0296] Step 3: Diethyl azodicarboxylate (0.166 mL, 1.05 mmol) was
added slowly to a solution of tert-butyl
2-(4-chlorophenyl)-2-hydroxyethyl(isopropyl)carbamate (265 mg,
0.843 mmol),
(R)-4-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)phenol
(159 mg, 0.703 mmol), and triphenylphosphine (276 mg, 1.05 mmol) in
tetrahydrofuran (7.0 mL) at -30.degree. C. (bath). The mixture was
allowed to warm up to room temperature and stirred overnight. Most
of the starting material remained. Triphenylphosphine (276 mg, 1.05
mmol) was added. The contents were cooled at -30.degree. C. Diethyl
azodicarboxylate (0.166 mL, 1.05 mmol) was added. The mixture was
stirred at room temperature for 4 hours. tert-Butyl
2-(4-chlorophenyl)-2-hydroxyethyl(isopropyl)carbamate (265 mg,
0.843 mmol) was added. The mixture was stirred at room temperature
overnight. EtOAC (10 mL) and water (10 mL) were added. The organic
layer was separated. The aqueous layer was extracted with EtOAc
(2.times.10 mL). The combined organic solutions were dried
(Na.sub.2SO.sub.4). The crude was purified by flash chromatography
to give (R)-tert-butyl
2-(4-chlorophenyl)-2-(4-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-
-yl)phenoxy)ethyl(isopropyl)carbamate (26 mg, 7%).
[0297] Step 4: Trifluoroacetic acid (0.5 mL) was added dropwise to
a solution of (R)-tert-butyl
2-(4-chlorophenyl)-2-(4-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-
-yl)phenoxy)ethyl(isopropyl)carbamate (26 mg, 0.0498 mmol) in
methylene chloride (1.0 mL) at 0.degree. C. The mixture was stirred
at 0.degree. C. for 10 minutes and then at room temperature for 2
hours. The contents were concentrated. The resulting residue was
purified by reverse-phase HPLC to give
(R)--N-(2-(4-chlorophenyl)-2-(4-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyr-
imidin-4-yl)phenoxy)ethyl)propan-2-amine as TFA salt (7.5 mg, 23%).
.sup.1H NMR (d6-DMSO, 500 MHz) .delta..quadrature.(ppm): 8.9 (s,
1H), 8.7 (s, br, 2H), 7.8 (t, 2H), 7.5 (m, 4H), 7.1 (d, 2H), 5.7
(d, 1H), 3.8 (m, 1H), 3.4-3.5 (m, 2H), 3.3-3.4 (m, 1H), 3.0-3.1 (m,
1H), 2.9 (m, 1H), 2.3 (m, 1H), 1.7 (m, 1H), 1.3 (dd, 6H), 0.9 (dd,
3H). MS (422.3, M+1).
Example 8
##STR00071##
[0298]
(5R,7R)-4-(3-(Amino(4-chlorophenyl)methyl)-5,6-dihydro-[1,2,4]triaz-
olo[4,3-a]pyrazin-7(8H)-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-
-7-ol
[0299] Step 1: 1,1-Carbonyldiimidazole (1.60 g, 9.89 mmol) was
added to a solution of
2-(tert-butoxycarbonylamino)-2-(4-chlorophenyl)acetic acid (2.57 g,
9.00 mmol) in CH.sub.2Cl.sub.2 (22.5 mL, 9.00 mmol), and the
mixture was stirred for 30 minutes until gas evolution ceased.
Hydrazine (0.367 mL, 11.7 mmol) was then added to this solution,
and the mixture was stirred at room temperature for 12 hours. The
reaction mixture was diluted with CH.sub.2Cl.sub.2 and washed with
NaHCO.sub.3 (10%). The organic layer was dried (Na.sub.2SO.sub.4)
and concentrated. The residue was purified by reversed-phase
chromatography (Biotage SP4, 40+M, C.sub.18, 25% to 100%
MeCN/H.sub.2O). Fractions containing product were poured into a
separatory funnel and extracted with ethyl acetate (3.times.). The
combined organic layers were dried and concentrated to give
tert-butyl 1-(4-chlorophenyl)-2-hydrazinyl-2-oxoethylcarbamate as a
solid (1.95 g, 72%). LCMS (APCI.sup.+) M+H.sup.+: 299 (5%);
M+H.sup.+-Boc: 199 (100%); rt=2.78 minutes. .sup.1H NMR (400 MHz,
CDCl.sub.3) .delta. 7.73 (s, 1H), 7.30 (s, 4H), 5.85 (d, J=6.6 Hz,
1H), 5.21 (s, 1H), 3.87 (s, 2H), 1.42 (s, 9H).
[0300] Step 2: Triethyloxonium hexafluorophosphate (3.20 g, 28.0
mmol) was added to a solution of tert-butyl
3-oxopiperazine-1-carboxylate (2.54 g, 12.7 mmol) in
CH.sub.2CL.sub.2 (31.7 mL) at 0.degree. C., and the resulting
solution was stirred at room temperature for 18 hours. The reaction
mixture was washed with saturated NaHCO.sub.3, dried
(Na.sub.2SO.sub.4), and concentrated to give tert-butyl
3-ethoxy-5,6-dihydropyrazine-1(2H)-carboxylate as an oil. LCMS
(APCI.sup.+) M+H-t-Bu: 173 (40%); rt=3.14 minutes.
[0301] Step 3: A solution of tert-butyl
3-ethoxy-5,6-dihydropyrazine-1(2H)-carboxylate (0.762 g, 3.34 mmol)
and tert-butyl 1-(4-chlorophenyl)-2-hydrazinyl-2-oxoethylcarbamate
(1.00 g, 3.34 mmol) in toluene (6.67 mL, 3.34 mmol) was heated to
reflux and stirred at this temperature for 1 hour. The reaction
mixture was cooled to room temperature, diluted with ethyl acetate,
and washed with water. The organic layer was dried and concentrated
to give a residue that was purified by reverse phase chromatography
(Biotage SP4, 40+M, C.sub.18, 10% to 100% ACN/H.sub.2O) to afford
tert-butyl
3-((tert-butoxycarbonylamino)(4-chlorophenyl)methyl)-5,6-dihydro-[1,2,4]t-
riazolo[4,3-a]pyrazine-7(8H)-carboxylate (0.92 g, 60%). LCMS
(APCI.sup.+) M+H.sup.+: 463 (25%), 464 (90%), 466 (25%), 467 (5%);
rt=3.56 minutes. .sup.1H NMR (400 MHz, CDCl.sub.3) .delta.
.quadrature.7.33 (d, J=8.6 Hz, 1H), 7.29 (d, J=8.6 Hz, 1H), 6.12
(d, J=7.8 Hz, 1H), 5.87 (d, J=7.8 Hz, 1H), 4.84 (d, J=17.6 Hz, 1H),
4.75 (d, J=17.6 Hz, 1H), 3.93 (m, 1H), 3.80 (m, 1H), 3.73 (m, 1H),
3.49 (m, 1H), 1.47 (s, 9H), 1.41 (s, 9H).
[0302] Step 4: A solution of tert-butyl
3-((tert-butoxycarbonylamino)(4-chlorophenyl)methyl)-5,6-dihydro-[1,2,4]t-
riazolo[4,3-a]pyrazine-7(8H)-carboxylate (0.652 g, 1.41 mmol) in
MeOH (7.03 mL, 1.41 mmol) was saturated with HCl (g). The reaction
mixture was stirred at room temperature for 30 minutes and then
concentrated in vacuo to afford
(4-chlorophenyl)(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyraz-
in-3-yl)methanamine bishydrochloride as a foam (0.47 g, 99%). LCMS
(APCI.sup.+) M+H.sup.+: 264 (50%), 266 (10%), 267 (3%); rt=1.76
minutes. .sup.1H NMR (400 MHz, d.sub.6-DMSO) .delta.
.quadrature.10.62 (s, 1H), 10.18 (s, 1H), 9.43 (s, 3H), 7.57 (d,
J=8.6 Hz, 2H), 7.53 (d, J=8.6 Hz, 2H), 6.13 (m, 1H), 4.51 (m, 2H),
4.44 (m, 1H), 3.56 (m, 3H).
[0303] Step 5: A solution of
(4-chlorophenyl)(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)me-
thanamine bishydrochloride (50 mg, 0.149 mmol),
(5R,7R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-yl
4-nitrobenzoate (49.8 mg, 0.149 mmol) and DIEA (0.130 mL, 0.745
mmol) in NMP (0.30 mL, 0.149 mmol) was heated at 100.degree. C. for
1 day. The crude reaction mixture was diluted with methanol and
filtered. The filtrate was purified (C18, 5-95% MeCN/H.sub.2O+1%
TFA). The fractions containing product were isolated by basifying
with 25% NaOH and extracting into CH.sub.2Cl.sub.2/isopropanol
(3:1). The organic layers were concentrated, and the residue was
diluted with methanol. The solution was saturated with HCl (g). The
solution was evaporated to give
(5R,7R)-4-(3-(Amino(4-chlorophenyl)methyl)-5,6-dihydro-[1,2,4]triazolo[4,-
3-a]pyrazin-7(8H)-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-ol
bishydrochloride (15.3 mg, 25%). LCMS (APCI+) rt=2.28 min. .sup.1H
NMR (400 MHz, d.sub.6-DMSO+3 drops D.sub.2O) .delta. 8.83 (d, J=7.8
Hz, 1H), 7.55 (s, 4H), 6.02 (d, J=8.6 Hz, 1H), 5.30 (m, 1H), 4.54
(m, 1H), 4.36 (m, 1H), 4.24 (m, 1H), 4.09 (m, 1H), 3.48 (m, 1H),
2.21 (m, 1H), 2.08 (m, 1H), 1.26 (m, 1H), 1.19 (m, 3H).
Example 9
##STR00072##
[0304]
(5R,7R)-4-(3-(1-(4-Chlorophenyl)-2-(isopropylamino)ethyl)-5,6-dihyd-
ro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)-5-methyl-6,7-dihydro-5H-cyclope-
nta[d]pyrimidin-7-ol
[0305] Step 1: Using the procedure from Example 8, Step 1,
3-(tert-butoxycarbonyl(isopropyl)amino)-2-(4-chlorophenyl)propanoic
acid (2.00 g, 5.85 mmol) was used to afford tert-butyl
2-(4-chlorophenyl)-3-hydrazinyl-3-oxopropyl(isopropyl)carbamate
(1.90 g, 5.34 mmol, 91%). LCMS (APCI+) M+H.sup.+: 356 (100%), 358
(50%), rt 3.50 minutes.
[0306] Step 2: Using the procedure from Example 8, Step 3,
tert-butyl
2-(4-chlorophenyl)-3-hydrazinyl-3-oxopropyl(isopropyl)carbamate
(1.90 g, 5.34 mmol) was used to afford tert-butyl
3-(2-(tert-butoxycarbonyl(isopropyl)amino)-1-(4-chlorophenyl)ethyl)-5,6-d-
ihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylate (1.66 g,
3.18 mmol, 60%). LCMS (APCI+) M+H.sup.+: 519 (66%), 520 (95%), rt
4.09 minutes.
[0307] Step 3: Using the procedure from Example 8, Step 4,
tert-butyl
3-(2-(tert-butoxycarbonyl(isopropyl)amino)-1-(4-chlorophenyl)ethyl)-5,6-d-
ihydro-[1,2,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylate (1.66 g,
3.18 mmol) was used to afford
N-(2-(4-chlorophenyl)-2-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-
-3-yl)ethyl)propan-2-amine bishydrochloride (0.65 g, 1.66 mmol,
52%). LCMS (APCI+) M+H.sup.+: 320 (100%), 322 (25%), rt 1.89
minutes.
[0308] Step 4: Using the procedure from Example 8, Step 5,
N-(2-(4-chlorophenyl)-2-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-
-3-yl)ethyl)propan-2-amine bishydrochloride (95 mg, 0.15 mmol) was
used to afford
(5R,7R)-4-(3-(1-(4-chlorophenyl)-2-(isopropylamino)ethyl)-5,6-dihy-
dro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)-5-methyl-6,7-dihydro-5H-cyclop-
enta[d]pyrimidin-7-ol bishydrochloride (20 mg, 0.042 mmol, 28%).
LCMS APCI+(M+H.sup.+): rt 2.20 minutes. HPLC purity at 254 nm 98%,
rt=1.82 minutes. .sup.1H NMR (400 MHz, d.sub.6-DMSO+2 drops
D.sub.2O) .delta. 8.81 (d, J=6.6 Hz, 1H), 7.47 (d, J=8.2 Hz, 2H),
7.40 (dd, J=8.6 Hz, 3.5 Hz, 2H), 5.48 (dd, J=29, 16 Hz, 1H), 5.32
(td, J=8.2, 2.0 Hz, 1H), 5.21 (dd, J=32, 17 Hz, 1H), 4.84 (q, J=7
Hz, 1H), 4.31 (m, 2H), 3.84 (m, 1H), 3.71 (m, 1H), 3.43 (m, 4H),
2.23 (dd, J=12, 8.2 Hz, 1H), 2.12 (m, 1H), 1.26 (d, J=6.6 Hz, 6H),
1.16 (t, J=7.5 Hz, 3H).
Example 10
##STR00073##
[0309]
(4-Chlorophenyl)(7-((R)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimi-
din-4-yl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)methanamin-
e
[0310] Using the procedure from Example 8, Step 5, using
(R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidine and
(4-chlorophenyl)(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)me-
thanamine bishydrochloride (100 mg, 0.30 mmol) to afford
(4-chlorophenyl)(7-((R)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4--
yl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazin-3-yl)methanamine
bishydrochloride (75 mg, 0.19 mmol, 64%): LCMS APCI+(M+H.sup.+):
396 (60%), 398 (20%), rt 2.48 min. HPLC purity at 254 nm >99%,
rt=1.68 min. .sup.1H NMR (400 MHz, d.sub.6-DMSO) .quadrature. 9.51
(s, 3H), 8.85 (d, J=9.4 Hz, 1H), 7.61 (d, J=7.0 Hz, 2H), 7.55 (d,
J=7.8 Hz, 2H), 6.08 (br d, J=9.0 Hz, 1H), 5.42 (dd, J=26 Hz, 16 Hz,
1H), 5.19 (dd, J=21 Hz, 18 Hz, 1H), 4.55 (m, 1H), 4.29 (m, 1H),
3.79 (m, 1H), 3.14 (m, 1H), 2.96 (m, 1H), 2.32 (m, 1H), 1.82 (m,
1H), 1.15 (t, J=6.8 Hz, 2H), 1.04 (d, J=6.2 Hz, 3H).
Example 11
##STR00074##
[0311]
(5R,7R)-4-(3-((R)-1-Amino-2-(4-chlorophenyl)ethyl)-5,6-dihydro-[1,2-
,4]triazolo[4,3-a]pyrazin-7(8H)-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]p-
yrimidin-7-ol
[0312] Step 1: Using the procedure from Example 8, Step 1,
(R)-2-(tert-butoxycarbonylamino)-3-(4-chlorophenyl)propanoic acid
(3.02 g, 10.0 mmol) was used to afford (R)-tert-butyl
3-(4-chlorophenyl)-1-hydrazinyl-1-oxopropan-2-ylcarbamate (3.16 g,
10.0 mmol, >99%). LCMS APCI+(M+H.sup.+): 314 (95%), 316 (40%),
rt 2.92 minutes.
[0313] Step 2: Using the procedure from Example 8, Steps 2 and 3,
(R)-tert-butyl
3-(4-chlorophenyl)-1-hydrazinyl-1-oxopropan-2-ylcarbamate (3.16 g,
10.0 mmol) was used to afford (R)-tert-butyl
3-(1-(tert-butoxycarbonylamino)-2-(4-chlorophenyl)ethyl)-5,6-dihydro-[1,2-
,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylate (4.03 g, 8.43 mmol,
84%). LCMS APCI+(M+H.sup.+): 477 (20%), 478 (80%), 479 (20%), rt
3.63 minutes.
[0314] Step 3: Using the procedure from Example 8, Step 4,
(R)-tert-butyl
3-(1-(tert-butoxycarbonylamino)-2-(4-chlorophenyl)ethyl)-5,6-dihydro-[1,2-
,4]triazolo[4,3-a]pyrazine-7(8H)-carboxylate (4.03 g, 8.43 mmol)
was used to afford
(R)-2-(4-chlorophenyl)-1-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,-
3-a]pyrazin-3-yl)ethanamine bishydrochloride (2.47 g, 7.06 mmol,
84%). LCMS APCI+(M+H.sup.+): 278 (100%), rt 1.91 minutes. .sup.1H
NMR (400 MHz, d.sub.6-DMSO) .delta. 10.57 (br s, 1H), 10.43 (br s,
1H), 9.05 (br s, 3H), 7.33 (d, J=8.6 Hz, 2H), 7.20 (d, J=8.6 Hz,
2H), 4.95 (br s, 1H), 4.52 (m, 3H), 3.92 (m, 1H), 3.51 (m, 2H),
3.38 (m, 2H).
[0315] Step 4: Using the procedure from Example 8, Step 5,
(R)-2-(4-chlorophenyl)-1-(5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazi-
n-3-yl)ethanamine bishydrochloride (66 mg, 0.114 mmol) was used to
afford
(5R,7R)-4-(3-((R)-1-amino-2-(4-chlorophenyl)ethyl)-5,6-dihydro-[1,2,4]tri-
azolo[4,3-a]pyrazin-7(8H)-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimid-
in-7-ol hydrochloride (49 mg, 0.046 mmol, 40%). LCMS
APCI+(M+H.sup.+): 426 (100%), 428 (20%), rt 2.17 minutes. HPLC
purity at 254 nm >99%, rt=1.73 minutes. .sup.1H NMR (400 MHz,
d.sub.6-DMSO) .delta. 8.98 (s, 3H), 8.85 (s, 1H), 7.30 (d, J=7.8
Hz, 2H), 7.15 (d, J=8.2 Hz, 2H), 5.30 (m, 1H), 5.23 (s, 2H), 4.85
(br s, 1H), 4.35 (m, 1H), 4.00 (m, 1H), 3.76 (m, 1H), 3.70 (s, 1H),
3.61 (s, 1H), 3.51 (m, 1H), 3.40 (m, 1H), 3.27 (m, 1H), 2.16 (m,
2H), 1.15 (d, J=7.0 Hz, 3H).
Example 12
##STR00075##
[0316]
(5R,7R)-4-(4-(1-(4-Chlorophenyl)-2-(isopropylamino)ethylamino)pheny-
l)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-ol
hydrochloride
[0317] Step 1: TMSCN (19.9 mL, 149 mmol) was added to a solution of
4-chlorobenzaldehyde (20.0 g, 142 mmol), 4-bromoaniline (25.1 g,
146 mmol), and sulfamic acid (0.691 g, 7.11 mmol) in MeOH (56.9
mL), and the reaction mixture was stirred at room temperature for 6
hours. The reaction mixture was filtered, and the filter cake was
washed with ethanol, affording
2-(4-bromophenylamino)-2-(4-chlorophenyl)acetonitrile as a solid
(38.0 g, 83%). .sup.1H NMR (400 MHz, d.sub.6-DMSO) .delta.
.quadrature.7.53 (d, J=8.2 Hz, 2H), 7.44 (d, J=8.6 Hz, 2H), 7.37
(d, J=9.0 Hz, 2H), 6.64 (d, J=9.0 Hz, 2H), 5.38 (d, J=9.0 Hz, 1H),
4.06 (d, J=9.0 Hz, 1H).
[0318] Step 2: Lithium aluminum hydride (52.9 mL, 52.9 mmol) in THF
(1.0M) was added to a solution of
2-(4-bromophenylamino)-2-(4-chlorophenyl)acetonitrile (20.0 g, 62.2
mmol) in THF (311 mL, 62.2 mmol) at -78.degree. C. The reaction was
allowed to stir at -78.degree. C. and gradually warmed to room
temperature over 4 hours. The reaction mixture was poured into 1N
HCl. The solution was then basified and extracted with ethyl
acetate (3.times.). The combined organic layers were washed with
brine, dried and concentrated to give an oil. This residue was
purified by column chromatography, eluting first with ethyl
acetate/hexanes (2:1) then 5% MeOH/CH.sub.2Cl.sub.2->10%
MeOH/CH.sub.2Cl.sub.2+1% NH.sub.4OH, affording
N1-(4-bromophenyl)-1-(4-chlorophenyl)ethane-1,2-diamine as an oil
(9.60 g, 49%). .sup.1H NMR (400 MHz, CD.sub.3OD) .delta.
.quadrature.7.38 (d, 2H), 7.34 (d, 2H), 7.15 (d, 2H), 6.53 (d, 2H),
4.59 (dd, J=9.8, 4.7 Hz, 1H), 3.14 (dd, J=12.9, 4.7 Hz, 1H), 3.05
(dd, J=12.9, 9.4 Hz, 1H).
[0319] Step 3: NaBH(OAc).sub.3 (4.88 g, 23.0 mmol) was added to a
solution of N1-(4-bromophenyl)-1-(4-chlorophenyl)ethane-1,2-diamine
(5.00 g, 15.4 mmol) and propan-2-one (1.24 mL, 16.9 mmol) in
1,2-dichloroethane (51.2 mL, 15.4 mmol), and the resulting solution
was stirred at room temperature for 12 hours. The reaction was
followed by LCMS. The reaction mixture was quenched with saturated
NaHCO.sub.3 and extracted with ethyl acetate (3.times.). The
combined organic layers were dried and concentrated to give the
desired product as an oil, which was not further purified (5.40 g,
95%). .sup.1H NMR (400 MHz, CDCl3) .delta. 7.38 (d, J=9.0 Hz, 2H),
7.34 (d, J=9.0 Hz, 2H), 7.15 (d, J=9.0 Hz, 2H), 6.53 (d, J=9.0 Hz,
2H), 4.59 (m, 1H), 3.10 (m, 2H).
[0320] Step 4: A mixture of
N1-(4-bromophenyl)-1-(4-chlorophenyl)-N2-isopropylethane-1,2-diamine
(0.556 g, 1.51 mmol),
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (0.576
g, 2.27 mmol), KOAc (0.594 g, 6.05 mmol) and DMSO (7.56 mL, 1.51
mmol) was degassed with nitrogen for 5 minutes.
PdCl.sub.2(dppf)*CH.sub.2Cl.sub.2 (0.062 g, 0.076 mmol) was then
added to this solution, and the reaction was heated at 80.degree.
C. overnight under nitrogen. After 1 day, additional
4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (1.15
g, 4.54 mmol), KOAc (1.19 g, 12.1 mmol) and
PdCl.sub.2(dppf)*CH.sub.2Cl.sub.2 (0.124 g, 0.152 mmol) was added,
and the reaction was heated at 80.degree. C. for an additional 24
hours. The reaction mixture was diluted with ethyl acetate to
precipitate inorganic salts. The slurry was filtered to remove the
salts, and the filtrate was concentrated and loaded directly onto a
C.sub.18 samplet (40+M) and eluted on a Biotage Horizon (10%-100%
ACN/H.sub.2O+1% iPA, 1 mM NH.sub.4OAc). The column fractions
containing product were poured into a separatory funnel, basified
with 1N NaOH and extracted with ethyl acetate. The combined organic
layers were dried with sodium sulfate and concentrated to afford
the product as a foam (0.220 g, 35%). LCMS APCI+(M+H.sup.+): 415
(25%), 417 (10%), rt 3.76 minutes.
[0321] Step 5:
1-(4-Chlorophenyl)-N2-isopropyl-N1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxabor-
olan-2-yl)phenyl)ethane-1,2-diamine (0.075 g, 0.181 mmol),
Pd(PPh.sub.3).sub.4 (0.017 g, 0.015 mmol), and 2N Na.sub.2CO.sub.3
(0.226 mL, 0.452 mmol) were added to a solution of
(5R,7R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-yl
4-nitrobenzoate (0.050 g, 0.151 mmol) in n-propanol (0.464 mL,
0.151 mmol). The suspension was degassed by bubbling nitrogen
through the solution. The dark suspension was heated at 90.degree.
C. for 14 hours. The mixture was cooled to room temperature and
then concentrated. The resulting residue was diluted with ethyl
acetate and filtered. The filtrate was washed with saturated
NaHCO.sub.3 and brine. The organic layer was dried and
concentrated. The resulting residue was purified by Biotage SP4
(C.sub.18, 25+0-100% ACN). The product-containing fractions were
collected and repurified by Gilson C.sub.18 (5-95 ACN/H.sub.2O+1%
TFA). Tubes containing product were identified by LCMS, collected
and evaporated. The material was partially dissolved in
CH.sub.2Cl.sub.2, and HCl (g) was bubbled through the mixture to
precipitate solid
(5R,7R)-4-(4-(1-(4-chlorophenyl)-2-(isopropylamino)ethylamino)phenyl)-5-m-
ethyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-ol hydrochloride
(8.10 mg, 12%). LCMS APCI+(M+H.sup.+): 437 (100%), 439 (30%), rt
2.55 minutes. HPLC purity at 254 nm >99%, rt=2.17 minutes.
.sup.1H NMR (400 MHz, d.sub.6-acetone) .delta. 8.88 (s, 1H), 7.81
(d, J=8.2 Hz, 1H), 7.78 (d, J=8.6 Hz, 1H), 7.59 (d, J=3.5 Hz, 1H),
7.58 (d, J=3.1 Hz, 1H), 7.40 (d, J=8.2 Hz, 2H), 6.65 (t, J=9.0 Hz,
2H), 5.26 (m, 1H), 5.15 (td, J=7.4, 4.3 Hz, 1H), 3.88 (m, 2H), 3.62
(br s, 2H), 3.50 (br s, 1H), 2.17 (m, 2H), 1.45 (m, 2H), 1.41 (d,
J=6.6 Hz, 6H), 1.10 (t, J=7.0 Hz, 3H), 0.88 (m, 1H).
Example 13
##STR00076##
[0322]
(R)--N-(2-aminoethyl)-N-(4-chlorobenzyl)-4-(5-methyl-6,7-dihydro-5H-
-cyclopenta[d]pyrimidin-4-yl)benzamide
[0323] Step 1: A solution of Na.sub.2CO.sub.3 (1.65 mL, 1M) was
added to a solution of
(R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidine (253
mg, 1.50 mmol) and 4-methoxycarbonylphenylboronic acid (297 mg,
1.65 mmol) in 1,4-dioxane (4.5 mL). The mixture was sparged with
N.sub.2 for 2 minutes. The catalyst Pd(dppf)Cl.sub.2 (98 mg, 0.12
mmol) was added in one portion. The reaction vial was sealed and
heated in microwave to 110.degree. C. for 30 minutes. Water (20 mL)
was added to the mixture and extracted with DCM (3.times.15 mL).
The combined organics were dried (Na.sub.2SO.sub.4), filtered and
concentrated. The crude product was purified by flash
chromatography (0-50% EtOAc/hexane gradient elution) to give
(R)-methyl
4-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)benzoate as
an oil (300 mg, 70%). .sup.1H NMR (CDCl.sub.3, 500 MHz) .delta.
9.07 (s, 1H), 8.16 (d, =8.5 Hz, 2H), 7.88 (d, J=8.5 Hz, 2H), 3.96
(s, 3H), 3.81-3.76 (m, 1H), 3.15-3.00 (m, 2H), 2.44-2.40 (m, 1H),
1.78-1.74 (m, 1H), 1.01 (d, J=6.5 Hz, 3H).
[0324] Step 2: A solution of LiOH (64 mg, 2.68 mmol) in H.sub.2O
(10 mL) was added to a solution of (R)-methyl
4-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)benzoate
(359 mg, 1.34 mmol) in THF (6 mL) at 0.degree. C. The mixture was
allowed to warm up to room temperature and stirred overnight. The
volatile solvent was removed in vacuo. The aqueous layer was
acidified with 1N HCl to a pH of 3. A solid precipitated, and was
collected by filtration, washed with ether, and dried in vacuo to
give
(R)-4-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)benzoic
acid as a solid (306 mg, 90%).
[0325] Step 3: DIPEA (37 .mu.L, 0.21 mmol) and
O-(benzotriazol-1yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (80 mg, 0.21 mmol) were added to a solution of
(R)-4-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)benzoic
acid (51 mg, 0.2 mmol) in DMF (1 mL). The mixture was heated to
70.degree. C. for 18 hours. The mixture was diluted with DCM (10
mL). Saturated NH.sub.4Cl (10 mL) was added. The layers were
separated. The aqueous layer was extracted with DCM (2.times.10
mL). The combined organics were dried (Na.sub.2SO.sub.4), filtered
and concentrated. The crude product was purified by flash
chromatography (0-100% EtOAc/hexane gradient elution) to give
(R)-tert-butyl
2-(N-(4-chlorobenzyl)-4-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-
-yl)benzamido)ethylcarbamate as an oil (49 mg, 31%).
[0326] Step 4: TFA (182 .mu.L, 2.36 mmol) was added to a solution
of (R)-tert-butyl
2-(N-(4-chlorobenzyl)-4-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-
-yl)benzamido)ethylcarbamate (49 mg, 0.094 mmol) in DCM (1 mL). The
mixture was stirred at room temperature for 1.5 hours. The mixture
was concentrated in vacuo. The crude product was purified by
reverse phase HPLC to give
(R)--N-(2-aminoethyl)-N-(4-chlorobenzyl)-4-(5-methyl-6,7-dihydro-5H-cyclo-
penta[d]pyrimidin-4-yl)benzamide as an oil (9 mg, 22%). MS (APCI+)
[M+H].sup.+ 421.2. .sup.1H NMR (D.sub.2O, 500 MHz) .delta. 9.09 (s,
1H), 7.81 (d, J=8.5 Hz, 2H), 7.63 (d, J=8.0 Hz, 2H), 7.35 (d, J=8.5
Hz, 2H), 7.15 (D, J=8.0 Hz, 2H), 4.63 (s, 2H), 3.87 (t, J=6.5 Hz,
2H), 3.82-3.79 (m, 1H), 3.26-3.11 (m, 4H), 2.63 (s, 2H), 2.50-2.42
(m, 1H), 1.86-1.81 (m, 1H), 0.86 (s, J=7.00 Hz, 3H).
Example 14
##STR00077##
[0327]
(R)--N-(2-aminoethyl)-N-(4-trifluoromethylbenzyl)-4-(5-methyl-6,7-d-
ihydro-5H-cyclopenta[d]pyrimidin-4-vl)benzamide
[0328]
(R)--N-(2-Aminoethyl)-N-(4-trifluoromethylbenzyl)-4-(5-methyl-6,7-d-
ihydro-5H-cyclopenta[d]pyrimidin-4-yl)benzamide was prepared in a
similar manner as
(R)--N-(2-aminoethyl)-N-(4-chlorobenzyl)-4-(5-methyl-6,7-dihydr-
o-5H-cyclopenta[d]pyrimidin-4-yl)benzamide. MS (APCI+) [M+H].sup.+
455.2. .sup.1H NMR (D.sub.2O, 500 MHz) .delta. 8.86 (s, 1H),
7.77-7.96 (m, 4H), 7.56 (d, J=8.0 Hz, 2H0, 7.35 (d, J=8.0 Hz, 2H),
4.75 (s, 2H), 3.84 (t, J=6.0 Hz, 2H), 3.70-3.67 (m, 1H), 3.84 (t,
J=6.0 Hz, 2H), 3.24-2.87 (m, 2H), 2.41-2.35 (m, 1H), 1.76-1.71 (m,
1H), 1.27-1.19 (m, 1H), 0.81 (d, J=7.0 Hz, 3H).
Example 15
##STR00078##
[0329]
(R)--N-(2-aminoethyl)-N-(4-bromolbenzyl)-4-(5-methyl-6,7-dihydro-5H-
-cyclopenta[d]pyrimidin-4-yl)benzamide
[0330]
(R)--N-(2-Aminoethyl)-N-(4-bromolbenzyl)-4-(5-methyl-6,7-dihydro-5H-
-cyclopenta[d]pyrimidin-4-yl)benzamide was prepared in a similar
manner as
(R)--N-(2-aminoethyl)-N-(4-chlorobenzyl)-4-(5-methyl-6,7-dihydro-5H-cyclo-
penta[d]pyrimidin-4-yl)benzamide. MS (APCI+) [M+H].sup.+ 465.2.
.sup.1H NMR (D.sub.2O, 500 MHz) .delta. 8.82 (s, 1H), 7.68 (d,
J=8.5 Hz, 2H), 7.52 (d, J=8.5 Hz, 2H), 7.48 (d, J=8.0 Hz, 7.06 (d,
J=8.0 Hz, 2H), 4.58 (s, 2H), 3.75 (t, J=6.5 Hz, 2H), 3.69-3.66 (m
1H), 3.22-2.78 (m, 4H), 2.37-2.29 (m, 1H), 1.71-1.67 (m, 1H),
1.22-1.37 (m, 2H), 0.75 (d, J=6.0 Hz, 3H).
Example 16
##STR00079##
[0331]
(S)-2-(4-bromophenyl)-3-(tert-butylamino)-1-(4-((5R,7R)-7-hydroxy-5-
-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)-5,6-dihydropiperidin-1-
(2H)-yl)propan-1-one
[0332] Step 1: A solution of Na.sub.2CO.sub.3 (2 mL, 2M) was added
to a solution of
(5R,7R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-yl
4-nitrobenzoate (334 mg, 1.00 mmol),
3,6-dihydro-2H-pyridine-1-tert-butoxycarbonyl-4-boronic acid,
pinacol ester (340 mg, 1.10 mmol) in 1,4-dioxane (6 mL). The
mixture was sparged with N.sub.2 for 2 minutes. The catalyst
Pd(dppf)Cl.sub.2 (65 mg, 0.08 mmol) was added in one portion. The
reaction vial was sealed and heated in microwave to 120.degree. C.
for 20 minutes. A solution of LiOH (0.7 mL, 3M) was added. The
mixture was stirred at room temperature for 18 hours. Water (30 mL)
was added to the mixture, extracted with DCM (3.times.20 mL). The
combined organics were dried (Na.sub.2SO.sub.4), filtered and
concentrated. The crude product was purified by flash
chromatography (0-5% MeOH/DCM gradient elution) to give tert-butyl
4-((5R,7R)-7-hydroxy-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)-
-5,6-dihydropyridine-1(2H)-carboxylate as an oil (221 mg, 67%).
.sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. 9.01 (s, 1H), 6.44 (s,
1H), 5.30 (m, 1H), 4.16-4.11 (m, 2H), 3.71-3.55 (m, 3H), 2.83-2.78
(m, 1H), 2.50-2.30 (m, 1H), 2.28-2.24 (m, 2H), 1.50 (s, 9H),
1.28-1.22 (m, 3H).
[0333] Step 2: TFA (193 .mu.L, 2.5 mmol) was added to a solution of
tert-butyl
4-((5R,7R)-7-hydroxy-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)-
-5,6-dihydropyridine-1(2H)-carboxylate (33 mg, 0.10 mmol) in DCM (1
mL). The mixture was stirred at room temperature for 1 hour, and
concentrated to give
(5R,7R)-5-methyl-4-(1,2,3,6-tetrahydropyridin-4-yl)-6,7-dihydro-5-
H-cyclopenta[d]pyrimidin-7-ol as an oil, which was used further
without purification.
[0334] Step 3: DIPEA (174 .mu.L, 1.0 mmol) and
O-(benzotriazol-1yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (42 mg, 0.11 mmol) were added to a solution of
(5R,7R)-5-methyl-4-(1,2,3,6-tetrahydropyridin-4-yl)-6,7-dihydro-5H-cyclop-
enta[d]pyrimidin-7-ol (23 mg, 0.10 mmol) and
(S)-2-(4-bromophenyl)-3-(tert-butylamino)propanoic acid (33 mg,
0.11 mmol) in DCM (1 mL). The mixture was stirred at room
temperature for 1 hour. The reaction was quenched with saturated
NH.sub.4Cl, extracted with DCM (2.times.10 mL). The combined
organics were dried (Na.sub.2SO.sub.4), filtered and concentrated.
The crude product was purified by reverse phase HPLC to give
(S)-2-(4-bromophenyl)-3-(tert-butylamino)-1-(4-((5R,7R)-7-hydroxy-5-methy-
l-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)-5,6-dihydropiperidin-1(2H)-y-
l)propan-1-one ditrifluoroacetic acid as a solid (13 mg, 24%). MS
(APCI+) [M+H].sup.+ 514.2. .sup.1H NMR (D.sub.2O, 400 MHz) 8.66 (s,
1H), 7.46 (d, J=8.4 Hz, 2H), 7.10 (J=8.4 Hz, 2H), 6.18-6.15 (m,
1H), 5.06-5.00 (m, 1H), 4.44-4.40 (m, 1H), 4.25-4.22 (m, 1H),
4.01-3.65 (m, 5H), 3.52-3.45 (m, 2H), 3.38-3.05 (m, 3H), 2.05-1.82
(m, 3H), 1.17 (s, 9H), 0.77 (d, J=7.2 Hz, 3H).
Example 17
##STR00080##
[0335]
(S)-2-(4-bromophenyl)-3-(tert-butylamino)-1-(4-((5R,7R)-7-hydroxy-5-
-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)piperidin-1-yl)propan-1-
-one
[0336] Step 1: 5% Pd/C (10 mg) was added to a solution of
tert-butyl
4-((5R,7R)-7-hydroxy-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)-
-5,6-dihydropyridine-1(2H)-carboxylate (17 mg, 0.05 mmol) in EtOAc
(1 mL). The mixture was stirred at room temperature under a
hydrogen atmosphere overnight. The mixture was filtered through
celite and concentrated to give tert-butyl
4-((5R,7R)-7-hydroxy-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)-
piperidine-1-carboxylate as an oil (13 mg, 78%), which was used
further without purification.
[0337] Step 2: TFA (77 .mu.L, 1.0 mmol) was added to a solution of
tert-butyl
4-((5R,7R)-7-hydroxy-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)-
piperidine-1-carboxylate (13 mg, 0.04 mmol) in DCM (1 mL). The
mixture was stirred at room temperature for 1 hour and concentrated
to give
(5R,7R)-5-methyl-4-(piperidin-4-yl)-6,7-dihydro-5H-cyclopenta[d]pyrimidin-
-7-ol as an oil, which was used further without purification.
[0338] Step 3: DIPEA (70 L, 0.40 mmol) and
O-(benzotriazol-1yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (16 mg, 0.042 mmol) were added to a solution of
(5R,7R)-5-methyl-4-(piperidin-4-yl)-6,7-dihydro-5H-cyclopenta[d]pyrimidin-
-7-ol (9.3 mg, 0.04 mmol) and
(S)-2-(4-bromophenyl)-3-(tert-butylamino)propanoic acid (12 mg, 0.4
mmol) in DCM (1 mL). The mixture was stirred at room temperature
for 1 hour. The reaction was quenched with sat. NH.sub.4Cl and
extracted with DCM (2.times.10 mL). The combined organics were
dried (Na.sub.2SO.sub.4), filtered and concentrated. The crude
product was purified by reverse phase HPLC to give
(S)-2-(4-bromophenyl)-3-(tert-butylamino)-1-(4-((5R,7R)-7-hydroxy-5-methy-
l-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)piperidin-1-yl)propan-1-one
ditrifluoroacetic acid as a solid (4.6 mg, 22%). MS (APCI+)
[M+H].sup.+ 516.2. .sup.1H NMR (D.sub.2O, 500 MHz) .delta. 8.85 (s,
1H), 7.68 (d, J=7.5 Hz, 2H), 7.34 (d, J=7.5 Hz, 2H), 5.33 (t, J=4.5
Hz, 1H), 4.64-4.59 (m, 1H), 4.38-4.34 (m, 1H), 3.97-3.89 (m, 1H),
3.70-3.47 (m, 2H), 3.38-3.13 (m, 3H), 2.91-2.82 (m, 2H), 2.35-2.29
(m, 1H), 2.14-2.07 (m, 1H), 1.94-1.77 (m, 2H), 1.58-1.47 (m, 1H),
1.40 (s, 9H), 1.18 (d, J=7.0 Hz, 3H).
Example 18
##STR00081##
[0339]
(S)-2-(4-chlorophenyl)-2-((S)-5,5-dimethylpyrrolidin-2-yl)-1-(4-((R-
)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)piperidin-1-yl)ethan-
one
[0340] Step 1: A solution of Na.sub.2CO.sub.3 (0.36 mL, 2M) was
added to a solution of
(R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidine (101
mg, 0.60 mmol),
3,6-dihydro-2H-pyridine-1-tert-butoxycarbonyl-4-boronic acid,
pinacol ester (204 mg, 0.66 mmol) in 1,4-dioxane (1.8 mL). The
mixture was sparged with N.sub.2 for 2 minutes. The catalyst
Pd(PPh.sub.3).sub.2Cl.sub.2 (21 mg, 0.03 mmol) was added in one
portion. The mixture was heated at 110.degree. C. under N.sub.2 for
8 hours. Water (20 mL) was added to the mixture and extracted with
DCM (3.times.15 mL). The combined organics were dried
(Na.sub.2SO.sub.4), filtered and concentrated. The crude product
was purified by flash chromatography (first with 0-50%
EtOAc/hexane, then with 0-4% MeOH/DCM gradient elution) to give
(R)-tert-butyl
4-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)-5,6-dihydropyridi-
ne-1(2H)-carboxylate as an oil (127 mg, 67%). .sup.1H NMR
(CDCl.sub.3, 500 MHz) .delta. 8.91 (s, 1H), 6.31 (s, 1H), 3.64-3.55
(m, 1H), 3.10-3.03 (m, 2H), 2.98-2.90 (m, 2H), 2.83-2.78 (m, 1H),
2.42-2.34 (m, 2H), 1.77-1.69 (m, 2H). 1.49 (s, 9H), 1.19 (d, J=7.0
Hz, 3H).
[0341] Step 2: 5% Pd/C (40 mg) was added to a solution of
(R)-tert-butyl
4-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)-5,6-dihydropyridi-
ne-1(2H)-carboxylate (63 mg, 0.20 mmol) in EtOAc (4 mL). The
mixture was stirred at room temperature under a hydrogen atmosphere
overnight. The mixture was filtered through celite and concentrated
to give (R)-tert-butyl
4-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)piperidine-1-carbo-
xylate as an oil (58 mg, 91%), which was used further without
purification.
[0342] Step 3: TFA (193 .mu.L, 2.50 mmol) was added to a solution
of (R)-tert-butyl
4-(5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)piperidine-1-carbo-
xylate (32 mg, 0.10 mmol) in DCM (1 mL). The mixture was stirred at
room temperature for 1 hour and concentrated to give
(R)-5-methyl-4-(piperidin-4-yl)-6,7-dihydro-5H-cyclopenta[d]pyrimidine
as an oil, which was used further without purification.
[0343] Step 4: DIPEA (174 .mu.L, 1.00 mmol) and
O-(benzotriazol-1yl)-N,N,N',N'-tetramethyluronium
hexafluorophosphate (25 mg, 0.065 mmol) were added to a solution of
(R)-5-methyl-4-(piperidin-4-yl)-6,7-dihydro-5H-cyclopenta[d]pyrimidine
(22 mg, 0.10 mmol) and
(S)-2-((S)-1-(tert-butoxycarbonyl)-5,5-dimethylpyrrolidin-2-yl)-2-(4-chlo-
rophenyl)acetic acid (20 mg, 0.054 mmol) in DCM (2 mL). The mixture
was stirred at room temperature for 2 hours. The reaction was
quenched with saturated NH.sub.4Cl and extracted with DCM
(2.times.10 mL). The combined organics were dried
(Na.sub.2SO.sub.4), filtered and concentrated. The crude product
was purified by flash chromatography (0-5% MeOH/DCM gradient
elution) to give (S)-tert-butyl
5-((S)-1-(4-chlorophenyl)-2-(4-((R)-5-methyl-6,7-dihydro-5H-cyclopenta[d]-
pyrimidin-4-yl)piperidin-1-yl)-2-oxoethyl)-2,2-dimethylpyrrolidine-1-carbo-
xylate as a solid (26 mg, 84%). .sup.1H NMR (CD.sub.3OD, 500 MHz)
.delta. 8.73 (s, 3H), 7.42 (d, J=8.5 Hz, 2H), 7.25 (d, J=8.5 Hz,
2H), 4.73-4.70 (m, 2H), 4.40-4.35 (m, 1H), 3.89-3.86 (m, 1H),
3.76-3.70 (m, 2H), 3.49-3.43 (m, 1H), 3.25-3.20 (m, 2H), 3.18-2.97
(m, 2H), 2.86-2.70 (m, 3H), 2.34-2.25 (m, 1H), 2.16-2.13 (m, 1H),
1.83-1.67 (m, 2H), 1.54 (s, 9H), 1.38 (s, 3H), 1.36 (s, 3H),
1.25-1.20 (m, 3H).
[0344] Step 5: A solution of 4M HCl in 1,4-dioxane (0.344 mL) was
added to a solution of (S)-tert-butyl
5-((S)-1-(4-chlorophenyl)-2-(4-((R)-5-methyl-6,7-dihydro-5H-cyclopenta[d]-
pyrimidin-4-yl)piperidin-1-yl)-2-oxoethyl)-2,2-dimethylpyrrolidine-1-carbo-
xylate (26 mg, 0.046 mmol) in 1,4-dioxane (1 mL) at 0.degree. C.
The mixture was allowed to warm up and stirred at room temperature
for 4 hours. The mixture was concentrated in vacuo. The resulting
residue was dissolved in minimal DCM and added to the ether (5 mL)
at 0.degree. C. A solid precipitated. The mixture was decanted and
dried in vacuo to give
(S)-2-(4-chlorophenyl)-2-((S)-5,5-dimethylpyrrolidin-2-yl)-1-(4-((R)-5-me-
thyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-4-yl)piperidin-1-yl)ethanone
dihydrochloride as a solid (26 mg, 100%). MS (APCI+) [M+H].sup.+
467.3. .sup.1H NMR (D.sub.2O, 500 MHz) .delta. 8.96 (s, 1H), 7.48
(d, J=8.5 Hz, 2H), 7.38 (d, J=8.5 Hz, 2H), 4.60-4.20 (m, 3H),
4.01-3.52 (m, 6H), 3.48-3.05 (m, 4H), 2.85-2.82 (m, 1H), 2.43-2.36
(m, 1H), 1.96-1.86 (m, 5H), 1.32 (s, 3H), 1.1.31 (s, 3H), 1.19 (d,
J=7.5 Hz, 3H).
Example 19
##STR00082##
[0345]
(S)-2-(4-chlorophenyl)-1-(4-((5R,7R)-7-hydroxy-5-methyl-6,7-dihydro-
-5H-cyclopenta[d]pyrimidin-4-yl)-1,4-diazepan-1-yl)-3-(isopropylamino)prop-
an-1-one
[0346] Step 1:
(5R,7R)-4-Chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-yl
4-nitrobenzoate (65 mg, 0.195 mmol) was dissolved in i-PrOH (5 mL),
and then tert-butyl 1,4-diazepane-1-carboxylate (51 mg, 0.253 mmol)
was added. N,N-Diisopropylethylamine (49 mg, 0.350 mmol) was added,
and the reaction mixture was heated to 80.degree. C. for 12 hours,
after which the solvents were removed under reduced pressure. The
resulting residue was taken up into EtOAc and then washed twice
with water and once with brine. The organic portion was dried over
magnesium sulfate, filtered, and then concentrated. The resulting
residue was purified via silica gel chromatography to give
tert-butyl
4-((5R,7R)-5-methyl-7-(4-nitrobenzoyloxy)-6,7-dihydro-5H-cyclopenta[d]pyr-
imidin-4-yl)azepane-1-carboxylate (78 mg, 81%). LCMS (APCI+)
M+=498.1, Rt=3.81 min.
[0347] Step 2: tert-Butyl
4-((5R,7R)-5-methyl-7-(4-nitrobenzoyloxy)-6,7-dihydro-5H-cyclopenta[d]pyr-
imidin-4-yl)azepane-1-carboxylate (78 mg, 0.160 mmol) was dissolved
in dichloromethane (2 mL) and then HCl (4M in dioxane, 0.58 mL) was
added. The resulting mixture was stirred at ambient for 4 hours, at
which time it was concentrated via rotary evaporation. The
resulting
(5R,7R)-4-(1,4-diazepan-1-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimi-
din-7-yl 4-nitrobenzoate hydrochloride (64 mg, 94%) was used
without further purification. LCMS (APCI+) M+H+=398.1, Rt=2.32
min.
[0348] Step 3:
(5R,7R)-4-(1,4-Diazepan-1-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimi-
din-7-yl 4-nitrobenzoate hydrochloride (61 mg, 0.140 mmol) and
3-(tert-butoxycarbonyl(isopropyl)amino)-2-(4-chlorophenyl)propanoic
acid (48 mg, 0.140 mmol) were suspended in dichloromethane (5 mL).
N,N-Diisopropylethylamine (54 mg, 0.420 mmol) was then added. HBTU
(53 mg, 0.140 mmol) was added, and the resultant solution was
stirred at ambient for 16 hours. After this time, the reaction
mixture was quenched by the addition of saturated sodium carbonate
solution and then extracted twice with dichloromethane. The
combined organic portions were dried over sodium sulfate, filtered,
and concentrated. The residue thus obtained was purified by silica
gel chromatography to give
(5R,7R)-4-(4-((S)-3-(tert-butoxycarbonyl(isopropyl)amino)-2-(4-chlorophen-
yl)propanoyl)-1,4-diazepan-1-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyri-
midin-7-yl 4-nitrobenzoate (50 mg, 50%). LCMS (APCI+) M+=721.1,
Rt=4.51 min.
[0349] Step 4:
(5R,7R)-4-(4-((S)-3-(tert-butoxycarbonyl(isopropyl)amino)-2-(4-chlorophen-
yl)propanoyl)-1,4-diazepan-1-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyri-
midin-7-yl 4-nitrobenzoate (50 mg, 0.069 mmol) was dissolved in a
THF/water mixture (1:1, 2 mL), and then solid lithium hydroxide (6
mg, 0.139 mmol) was added. The resultant mixture was stirred at
ambient for 16 hours, at which time it was diluted with EtOAc and
then washed twice with saturated sodium bicarbonate solution. The
organic layer was dried over magnesium sulfate, filtered and
concentrated. The resulting residue was taken up in dichloromethane
(5 mL), and then HCl (4M in dioxane, 0.25 mL) was added. The
mixture was stirred for 16 hours, at which time solvents were
removed via rotary evaporation. The resulting residue was dissolved
in dichloromethane (1 mL) and then added to diethyl ether (50 mL).
The resulting precipitate was collected by filtration and dried to
give
(S)-2-(4-chlorophenyl)-1-(4-((5R,7R)-7-hydroxy-5-methyl-6,7-dihydro--
5H-cyclopenta[d]pyrimidin-4-yl)-1,4-diazepan-1-yl)-3-(isopropylamino)propa-
n-1-one dihydrochloride (29 mg, 78%). LCMS (APCI+) M+=472.2,
Rt=2.05 min.
Example 20
##STR00083##
[0350]
(5R,7R)-4-(3-(1-(4-chlorophenyl)-2-(isopropylamino)ethyl)benzo[d]is-
othiazol-6-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-ol
[0351] Step 1: Oxalyl chloride (3.7 mL, 42 mmol) was added dropwise
to a solution of 3-bromothiophenol (5.00 g, 26.4 mmol) in ether (20
mL). The mixture was heated at reflux for 1.5 hours, cooled to room
temperature, and concentrated in vacuo. The resulting residue was
taken up in DCM (50 mL) and cooled to 0.degree. C. Aluminum
chloride (4.23 g, 31.7 mmol) was added in portions. The resultant
mixture was stirred at reflux for 30 minutes, cooled to room
temperature and poured into ice water with stirring. The organic
layer was separated and successively washed with saturated aqueous
NaHCO.sub.3, water and brine. The organic layer was dried and
concentrated in vacuo to give a solid, which was suspended in 20%
EtOAc in hexanes (50 mL) and heated at reflux for 10 minutes. After
cooling, the precipitated solid was collected by filtration to
afford crude 6-bromobenzo[b]thiophene-2,3-dione (3.22 g, 50%). The
6-bromobenzo[b]thiophene-2,3-dione was added to ammonium hydroxide
(35% aqueous solution, 40 mL) at 5-10.degree. C., followed by
dropwise addition of hydrogen peroxide (35% aqueous solution, 5.5
mL, 66 mmol). The resulting mixture was stirred at room temperature
for 30 minutes, and then filtered to give
6-bromobenzo[b]thiophene-3-carboxamide (1.30 g, 38%) as a solid.
.sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. .quadrature.8.66 (d,
J=8.8 Hz, 1H), 8.62 (s, 1H), 8.23 (s, 1H), 7.81 (s, 1H), 7.73 (d,
J=8.8 Hz, 1H).
[0352] Step 2: 10N NaOH solution (10 mL, 100 mmol) was added to a
solution of 6-bromobenzo[b]thiophene-3-carboxamide (1.20 g, 4.67
mmol) in MeOH (80 mL). The mixture was heated at reflux overnight.
After cooling, the mixture was acidified with 2N HCl. The resulting
precipitate was filtered and dried to give
6-bromobenzo[d]isothiazole-3-carboxylic acid (1.10 g, 91%) as a
solid. .sup.1H NMR (DMSO-d.sub.6, 400 MHz) .delta. .quadrature.8.65
(s, 1H), 8.55 (d, J=8.8 Hz, 1H), 7.77 (d, J=8.8 Hz, 1H).
[0353] Step 3: DIEA (11.7 mL, 67.4 mmol) and HBTU (7.03 g, 18.5
mmol) were added to a solution of
6-bromobenzo[d]isothiazole-3-carboxylic acid (4.35 g, 16.9 mmol)
and N,O-dimethylhydroxylamine hydrochloride (2.14 g, 21.9 mmol) in
DMF (100 mL). The reaction was stirred at room temperature for 2
hours. The mixture was partitioned between water and EtOAc. The
organic layer was washed with aqueous NaHCO.sub.3 solution and
brine, dried and concentrated. The residue was purified by column
chromatography (hexane:EtOAc, 3:1) to give
6-bromo-N-methoxy-N-methylbenzo[d]isothiazole-3-carboxamide (4.60
g, 91%) as a solid. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
.quadrature.8.12 (m, 2H), 8.59 (d, J=8.8 Hz, 1H), 3.83 (s, 3H),
3.49 (s, 3H).
[0354] Step 4: 4-Chlorophenyl magnesium bromide (1.0N in THF, 16
mL, 16 mmol) was added to a stirred solution of
6-bromo-N-methoxy-N-methylbenzo[d]isothiazole-3-carboxamide (3.5 g,
12 mmol) in THF (100 mL). The reaction mixture was stirred at
0.degree. C. for 1 hour. The reaction was poured into 1N HCl and
extracted into ether. The combined organic layers were washed with
brine, dried and concentrated. The crude product was suspended in
ether and stirred for 15 minutes. The solid was collected by
filtration to give
(6-bromobenzo[d]isothiazol-3-yl)(4-chlorophenyl)methanone (3.3 g,
81%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta. .quadrature.8.60
(d, J=8.8 Hz, 1H), 8.23 (d, J=8.4 Hz, 2H), 8.19 (s, 1H), 7.68 (d,
J=8.8 Hz, 1H), 7.51 (d, J=8.4 Hz, 2H).
[0355] Step 5: A mixture of NaH (60% mineral oil dispersion, 0.060
g, 1.5 mmol) and DMSO (3 mL) was stirred at 70.degree. C. for 45
minutes. The solution was then cooled with cold water, and
methyltriphenylphosphonium bromide (0.58 g, 1.6 mmol) in DMSO (3
mL) was added dropwise. Stirring was continued for 15 minutes.
6-Bromobenzo[d]isothiazol-3-yl)(4-chlorophenyl)methanone (0.300 g,
0.850 mmol) was then added in a single portion. The mixture was
stirred at room temperature for 1.5 hours and then poured into
ice-water. The mixture was extracted with EtOAc. The combined
organic layers were washed with brine, dried and concentrated. The
resulting residue was purified by column chromatography
(hexane:DCM, 10:1 to 6:1) to give
6-bromo-3-(1-(4-chlorophenyl)vinyl)benzo[d]isothiazole (0.26 g,
87%) as a solid. .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
.quadrature.8.22 (d, J=8.8 Hz, 1H), 8.12 (s, 1H), 7.51 (d, J=8.8
Hz, 1H), 7.30 (m, 4H), 5.97 (s, 1H), 5.81 (s, 1H).
[0356] Step 6: A mixture of
6-bromo-3-(1-(4-chlorophenyl)vinyl)benzo[d]isothiazole (600 mg,
1.71 mmol), DMF (3 mL) and allylamine (3 mL) was stirred at room
temperature for 3 days. The reaction was partitioned between EtOAc
and water. The organic phase was washed with brine, dried and
concentrated. The residue was purified by column chromatography
(DCM:MeOH, 20:1) to give
N-(2-(6-bromobenzo[d]isothiazol-3-yl)-2-(4-chlorophenyl)ethyl)prop-2-en-1-
-amine (464 mg, 66%). .sup.1H NMR (CDCl.sub.3, 400 MHz) .delta.
.quadrature.8.06 (d, J=1.6 Hz, 1H), 7.59 (d, J=8.8 Hz, 1H), 7.41
(dd, J=8.8 Hz, J=1.6 Hz, 1H), 7.25 (m, 4H), 5.86 (m, 1H), 5.16 (dd,
J=17.2 Hz, J=1.6 Hz, 1H), 5.08 (d, J=10.4 Hz, 1H), 4.72 (t, J=7.2
Hz, 1H), 3.62 (dd, J=12.0 Hz, J=8.0 Hz, 1H), 3.31 (m, 2H), 3.20
(dd, J=12.0 Hz, J=6.8 Hz, 1H).
[0357] Step 7: A mixture of
N-(2-(6-bromobenzo[d]isothiazol-3-yl)-2-(4-chlorophenyl)ethyl)prop-2-en-1-
-amine (0.441 g, 1.08 mmol),
1,3-dimethylpyrimidine-2,4,6(1H,3H,5H)-trione (0.507 g, 3.24 mmol),
Pd(PPh.sub.3).sub.4 (0.013 g, 0.011 mmol) and DCM (4 mL) was heated
at 35.degree. C. under N.sub.2 for 4 hours. After cooling, DCM was
evaporated. The resulting residue was taken up in ether, washed
with saturated NaHCO.sub.3 and brine, dried and concentrated. The
crude product was dissovled in THF (8 mL). Boc.sub.2O (0.28 g, 1.3
mmol) and Et.sub.3N (0.23 mL, 1.6 mmol) were added. The mixture was
stirred at room temperature for 1 hour. The solvent was evaporated,
and the residue partitioned between EtOAc and water. The organic
phase was separated and washed with brine, dried and concentrated.
The resulting residue was purified by column chromatography
(hexanes:EtOAc, 6:1) to give tert-butyl
2-(6-bromobenzo[d]isothiazol-3-yl)-2-(4-chlorophenyl)ethylcarbamate
(0.39 g, 77%) as a solid. LCMS (APCI+) m/z 467, 469 [M+H]+; Rt=3.71
min.
[0358] Step 8: Bis(pinacolato)diboron (0.25 g, 1.0 mmol),
tert-butyl
2-(6-bromobenzo[d]isothiazol-3-yl)-2-(4-chlorophenyl)ethylcarbamate
(0.39 g, 0.83 mmol) and potassium acetate (0.25 g, 2.5 mmol) were
added to DMF (4 mL). The reaction solution was deoxygenated and
then dichloro[1,1'-bis(diphenylphosphino)-ferrocene]palladium(II)
dichloromethane adduct (34 mg, 0.042 mmol) was added. The mixture
was heated to 80.degree. C. for 4 hours. The mixture was cooled to
room temperature and partitioned between EtOAc and water. The
aqueous phase was extrated with EtOAc. The combined organic layers
were washed with brine, dried and concentrated. The resulting
residue was purified by column chromatography (hexanes:EtOAc, 8:1)
to give tert-butyl
2-(4-chlorophenyl)-2-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benz-
o[d]isothiazol-3-yl)ethylcarbamate (0.36 g, 84%) as a solid. LCMS
(APCI+) m/z 515, 517 [M+H]+; Rt=4.96 min.
[0359] Step 9: DMF (3 mL) and 2M aqueous Na.sub.2CO.sub.3 (0.38 mL,
0.76 mmol) were added to a nitrogen flushed flask containing
tert-butyl
2-(4-chlorophenyl)-2-(6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benz-
o[d]isothiazol-3-yl)ethylcarbamate (150 mg, 0.291 mmol),
(5R,7R)-4-chloro-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-yl
4-nitrobenzoate (117 mg, 0.350 mmol) and
dichloro[1,1'-bis(diphenylphosphino)-ferrocene]palladium(II)
dichloromethane adduct (12 mg, 0.015 mmol). The mixture was heated
at 80.degree. C. for 1 hour. The mixture was cooled to room
temperature and partitioned between EtOAc and water. The combined
organic layers were washed with saturated aqueous NaHCO.sub.3 and
brine, dried and concentrated. The residue was purified by flash
chromatography on silica gel, eluting with hexanes:EtOAc (4:1 to
1:1) to give
(5R,7R)-4-(3-(2-(tert-butoxycarbonylamino)-1-(4-chlorophenyl)ethyl)benzo[-
d]isothiazol-6-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-yl
4-nitrobenzoate (102 mg, 51%) as an oil. LCMS (APCI+) m/z 630, 632
[M+H]+; Rt=4.68 min.
[0360] Step 10: 1N LiOH aqueous solution (0.30 mL, 0.30 mmol) was
added to a stirred solution of
(5R,7R)-4-(3-(2-(tert-butoxycarbonylamino)-1-(4-chlorophenyl)ethyl)benzo[-
d]isothiazol-6-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-yl
4-nitrobenzoate (102 mg, 0.149 mmol) in THF (3 mL). The reaction
mixture was stirred at room temperature overnight. The solvent was
evaporated. The resulting residue was partitioned between EtOAc and
water. The aqueous layer was extracted with EtOAc. The combined
organic layers were washed with brine, dried and concentrated. The
resulting residue was purified by column chromatography (DCM:MeOH,
60:1) to give tert-butyl
2-(4-chlorophenyl)-2-(6-((5R,7R)-7-hydroxy-5-methyl-6,7-dihydro-5H-cyclop-
enta[d]pyrimidin-4-yl)benzo[d]isothiazol-3-yl)ethylcarbamate (62
mg, 78%) as an oil. LCMS (APCI+) m/z 537, 539 [M+H]+; Rt=3.99
min.
[0361] Step 11: A solution of tert-butyl
2-(4-chlorophenyl)-2-(6-((5R,7R)-7-hydroxy-5-methyl-6,7-dihydro-5H-cyclop-
enta[d]pyrimidin-4-yl)benzo[d]isothiazol-3-yl)ethylcarbamate (41
mg, 0.076 mmol) in DCM was treated with 4N HCl in dioxane (0.5 mL).
The reaction was stirred at room temperature overnight. The
reaction was concentrated and triturated with ether (twice) to
yield
(5R,7R)-4-(3-(2-amino-1-(4-chlorophenyl)ethyl)benzo[d]isothiazol-6-yl)-5--
methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-ol trihydrochloride
(32 mg, 77%). LCMS (APCI+) m/z 437, 439 [M+H]+; Rt=2.48 min.
[0362] Step 12: DIEA (0.035 mL, 0.20 mmmol) was added to a stirred
suspension of
(5R,7R)-4-(3-(2-amino-1-(4-chlorophenyl)ethyl)benzo[d]isothiazol-6-yl)-5--
methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-ol trihydrochloride
(22 mg, 0.050 mmol) in DCE (1.5 mL). The suspension was shaken
until dissolved. A solution of acetone (0.022 mL, 0.30 mmol) in THF
(0.3 mL) was added. The reaction was allowed to stir at room
temperature for 15 minutes, at which point Na(OAc).sub.3BH (27 mg,
0.13 mmol) was added. The reaction was allowed to stir at room
temperature for 1 hour. The reaction mixture was diluted with DCM,
washed with saturated aqueous NaHCO.sub.3 solution and brine, dried
and concentrated. The resulting residue was purified by column
chromatography (DCM:7N ammonia in MeOH, 30:1) to give the free
base, which was taken up in DCM and acidified with 2N HCl in ether.
Removal of the solvents under reduced pressure gave
(5R,7R)-4-(3-(1-(4-chlorophenyl)-2-(isopropylamino)ethyl)benzo[d]isothiaz-
ol-6-yl)-5-methyl-6,7-dihydro-5H-cyclopenta[d]pyrimidin-7-ol
trihydrochloride. LCMS (APCI+) m/z 479, 481 [M+H]+; Rt=2.69
min.
[0363] While the invention has been described in conjunction with
the enumerated embodiments, it will be understood that they are not
intended to limit the invention to those embodiments. On the
contrary, the invention is intended to cover all alternatives,
modifications and equivalents, which may be included within the
scope of the present invention as defined by the claims. Thus, the
foregoing description is considered as illustrative only of the
principles of the invention.
[0364] The words "comprise," "comprising," "include," "including,"
and "includes" when used in this specification and in the following
claims are intended to specify the presence of stated features,
integers, components, or steps, but they do not preclude the
presence or addition of one or more other features, integers,
components, steps, or groups thereof.
Sequence CWU 1
1
1111PRTArtificial SequenceDescription of Artificial Sequence
Synthetic peptide 1Gly Arg Pro Arg Thr Ser Ser Phe Ala Glu Gly 1 5
10
* * * * *